8-substituted benzoazepines as toll-like receptor modulators

ABSTRACT

Provided are compositions and methods useful for modulation of signaling through the Toll-like receptors TLR7 and/or TLR8. The compositions and methods have use in the treatment of autoimmunity, inflammation allergy, asthma, graft rejection, graft versus host disease, infection, sepsis, cancer and immunodeficiency.

This application claims the benefit under 35 U.S.C. §119 of U.S.provisional patent application No. 60/710,004, filed Aug. 19, 2005, thecontents of which are incorporated by reference in their entirety.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention relates to methods and compositions for modulating immunefunction. More specifically, this invention relates to compositions andmethods for modulating TLR7- and/or TLR8-mediated signaling.

2. Description of the State of the Art

Stimulation of the immune system, which includes stimulation of eitheror both innate immunity and adaptive immunity, is a complex phenomenonthat can result in either protective or adverse physiologic outcomes forthe host. In recent years there has been increased interest in themechanisms underlying innate immunity, which is believed to initiate andsupport adaptive immunity. This interest has been fueled in part by therecent discovery of a family of highly conserved pattern recognitionreceptor proteins known as Toll-like receptors (TLRs) believed to beinvolved in innate immunity as receptors for pathogen-associatedmolecular patterns (PAMPs). Compositions and methods useful formodulating innate immunity are therefore of great interest, as they mayaffect therapeutic approaches to conditions involving autoimmunity,inflammation, allergy, asthma, graft rejection, graft versus hostdisease (GvHD), infection, cancer, and immunodeficiency.

Toll-like receptors (TLRs) are type I transmembrane proteins that alloworganisms (including mammals) to detect microbes and initiate an innateimmune response (Beutler, B., Nature 2004, 430:257-263). They containhomologous cytoplasmic domains and leucine-rich extracellular domainsand typically form homodimers that sense extracellular (or internalized)signals and subsequently initiate a signal transduction cascade viaadaptor molecules such as MyD88 (myeloid differentiation factor 88).There is such high homology in the cytoplasmic domains of the TLRs that,initially, it was suggested that similar signaling pathways exist forall TLRs (Re, F., Strominger, J. L., Immunobiology 2004, 209:191-198).Indeed, all TLRs can activate NF-kB and MAP kinases; however, thecytokine/chemokine release profiles derived from TLR activation appearsunique to each TLR. Additionally, the signaling pathway that TLRsstimulate is very similar to the pathway that the cytokine receptorIL-1R induces. This may be due to the homology that these receptorsshare, i.e., TIR (Toll/IL-1R homology) domains. Once the TIR domain isactivated in TLRs and MyD88 is recruited, activation of the IRAK familyof serine/threonine kinases results which eventually promotes thedegradation of Ik-B and activation of NF-kB (Means T. K., et al. LifeSci. 2000, 68:241-258). While it appears that this cascade is designedto allow extracellular stimuli to promote intracellular events, there isevidence that some TLRs migrate to endosomes where signaling can also beinitiated. This process may allow for intimate contact with engulfedmicrobes and fits with the role that these receptors play in the innateimmune response (Underhill, D. M., et al., Nature 1999, 401:811-815).This process might also allow host nucleic acids, released by damagedtissues (for example, in inflammatory disease) or apoptosis to trigger aresponse via endosomal presentation. Among mammals, there are 11 TLRsthat coordinate this rapid response. A hypothesis put forward years ago(Janeway, C. A., Jr., Cold Spring Harb. Symp. Quant. Biol. 1989,54:1-13) that the innate immune response initiates the adaptive immuneresponse through the pattern of TLR activation caused by microbes hasnow been substantiated. Thus, the pathogen-associated molecular patterns(PAMPs) presented by a diverse group of infectious organisms results ina innate immune response involving certain cytokines, chemokines andgrowth factors followed by a precise adaptive immune response tailoredto the infectious pathogen via antigen presentation resulting inantibody production and cytotoxic T cell generation.

Gram-negative bacterial lipopolysaccharide (LPS) has long beenappreciated as an adjuvant and immune-stimulant and as a pharmacologicaltool for inducing an inflammatory reaction in mammals similar to septicshock. Using a genetic approach, TLR4 was identified as the receptor forLPS. The discovery that LPS is an agonist of TLR4 illustrates theusefulness of TLR modulation for vaccine and human disease therapy(Aderem, A.; Ulevitch, R. J., Nature 2000, 406:782-787). It is nowappreciated that various TLR agonists can activate B cells, neutrophils,mast cells, eosinophils, endothelial cells and several types ofepithelia in addition to regulating proliferation and apoptosis ofcertain cell types.

To date, TLR7 and TLR8, which are somewhat similar, have beencharacterized as receptors for single-stranded RNA found in endosomalcompartments and thus thought to be important for the immune response toviral challenge. Imiquimod, an approved topical anti-viral/anti-cancerdrug, has recently been described as a TLR7 agonist that hasdemonstrated clinical efficacy in certain skin disorders (Miller R. L.,et al., Int. J. Immunopharm. 1999, 21:1-14). This small molecule drughas been described as a structural mimetic of ssRNA. TLR8 was firstdescribed in 2000 (Du, X., et al., European Cytokine Network 2000(September), 11(3):362-371) and was rapidly ascribed to being involvedwith the innate immune response to viral infection (Miettinen, M., etal., Genes and Immunity 2001 (October), 2(6):349-355).

Recently it was reported that certain imidazoquinoline compounds havingantiviral activity are ligands of TLR7 and TLR8 (Hemmi H., et al. (2002)Nat. Immunol. 3:196-200; Jurk M., et al. (2002) Nat. Immunol. 3:499).Imidazoquinolines are potent synthetic activators of immune cells withantiviral and antitumor properties. Using macrophages from wildtype andMyD88-deficient mice, Hemmi et al. recently reported that twoimidazoquinolines, imiquimod and resiquimod (R848), induce tumornecrosis factor (TNF) and interleukin-12 (IL-12) and activate NF-κB onlyin wildtype cells, consistent with activation through a TLR (Hemmi H.,et al. (2002) Nat. Immunol. 3:196-200). Macrophages from mice deficientin TLR7 but not other TLRs produced no detectable cytokines in responseto these imidazoquinolines. In addition, the imidazoquinolines induceddose-dependent proliferation of splenic B cells and the activation ofintracellular signaling cascades in cells from wildtype but not TLR7−/−mice. Luciferase analysis established that expression of human TLR7, butnot TLR2 or TLR4, in human embryonic kidney cells results in NF-κBactivation in response to resiquimod. The findings of Hemmi et al. thussuggest that these imidazoquinoline compounds are non-natural ligands ofTLR7 that can induce signaling through TLR7. Recently it was reportedthat R848 is also a ligand for human TLR8 (Jurk M., et al. (2002) Nat.Immunol. 3:499).

SUMMARY OF THE INVENTION

The compositions described herein are useful for modulating immuneresponses in vitro and in vivo. Such compositions will find use in anumber of clinical applications, such as in methods for treatingconditions involving unwanted immune activity, including inflammatoryand autoimmune disorders.

More specifically, one aspect of this invention provides a compound ofFormula I

and metabolites, solvates, tautomers, and pharmaceutically acceptablesalts and prodrugs thereof, wherein Y, R¹, R², R³, R⁴, R^(5a), R^(5b),and R^(5c) are as defined hereinbelow.

The invention also relates to pharmaceutical compositions comprising acompound of Formula I or a solvate, pharmaceutically acceptable prodrug,metabolite, or pharmaceutically acceptable salt thereof.

The inventive compounds may be used advantageously in combination withother known therapeutic agents. Accordingly, this invention also relatesto pharmaceutical compositions comprising a therapeutically effectiveamount of a compound of Formula I or a solvate, pharmaceuticallyacceptable prodrug, metabolite, or pharmaceutically acceptable saltthereof, in combination with a second therapeutic agent.

This invention further provides methods of modulating TLR7- and/orTLR8-mediated signaling, comprising contacting a cell expressing TLR7and/or TLR8 with an effective amount of a compound of Formula I, or asolvate, pharmaceutically acceptable prodrug, metabolite, orpharmaceutically acceptable salt thereof. In one aspect, the methodinhibits TLR7- and/or TLR8-mediated immunostimulatory signaling.

This invention further provides methods of modulating TLR7- and/orTLR8-mediated immunostimulation in a subject, comprising administeringto a patient having or at risk of developing TLR7- and/or TLR8-mediatedimmunostimulation a compound of Formula I, or a solvate,pharmaceutically acceptable prodrug, metabolite, or pharmaceuticallyacceptable salt thereof, in an amount effective to inhibit or promoteTLR7- and/or TLR8-mediated immunostimulation in the subject.

This invention further provides methods of treating a condition ordisorder treatable by modulation of TLR7- and/or TLR8-mediated cellularactivities, comprising administering to a warm-blooded animal, such as amammal, for example a human, having or at risk of developing saidcondition or disorder a compound of Formula I, or a solvate, metabolite,pharmaceutically acceptable prodrug, or pharmaceutically acceptable saltthereof, in an amount effective to treat said condition or disorder.

This invention further provides methods of modulating the immune systemof a mammal, comprising administering to a mammal a compound of FormulaI, or a solvate, metabolite, pharmaceutically acceptable prodrug, orpharmaceutically acceptable salt thereof, in an amount effective tomodulate said immune system.

Further provided is a compound of Formula I, or a solvate,pharmaceutically acceptable prodrug, or pharmaceutically acceptable saltthereof for use as a medicament in the treatment of the diseases orconditions described herein in a mammal, for example, a human, sufferingfrom such disease or condition. Also provided is the use of a compoundof Formula I, or a solvate, metabolite, pharmaceutically acceptableprodrug, or pharmaceutically acceptable salt thereof, in the preparationof a medicament for the treatment of the diseases and conditionsdescribed herein in a mammal, for example a human, suffering from suchdisorder.

This invention further provides kits comprising one or more compounds ofFormula I, or a solvate, metabolite, pharmaceutically acceptableprodrug, or pharmaceutically acceptable salt thereof. The kit mayfurther comprise a second compound or formulation comprising a secondpharmaceutical agent.

Additional advantages and novel features of this invention shall be setforth in part in the description that follows, and in part will becomeapparent to those skilled in the art upon examination of the followingspecification or may be learned by the practice of the invention. Theadvantages of the invention may be realized and attained by means of theinstrumentalities, combinations, compositions, and methods particularlypointed out in the appended claims.

BRIEF DESCRIPTION OF THE FIGURES

The accompanying drawings, which are incorporated herein and form a partof the specification, illustrate non-limiting embodiments of the presentinvention, and together with the description, serve to explain theprinciples of the invention.

In the Figures:

FIG. 1 shows a reaction scheme for the synthesis of compound 7.

FIG. 2 shows a reaction scheme for the synthesis of compound 8.

FIG. 3 shows a reaction scheme for the synthesis of compound 12.

FIG. 4 shows a reaction scheme for the synthesis of compound 24.

DETAILED DESCRIPTION OF THE INVENTION

In certain aspects, the invention provides compositions and methodsuseful for modulating TLR7- and/or TLR8-mediated signaling. Morespecifically, one aspect of this invention provides a compound ofFormula I

and metabolites, solvates, tautomers, and pharmaceutically acceptablesalts and prodrugs thereof, wherein:

Y is CF₂CF₃, CF₂CF₂R⁶, or an aryl or heteroaryl ring, wherein said aryland heteroaryl rings are substituted with one or more groupsindependently selected from alkenyl, alkynyl, Br, CN, OH, NR⁶R⁷,C(═O)R⁸, NR⁶SO₂R⁷, (C₁-C₆ alkyl)amino, R⁶OC(═O)CH═CH₂—, SR⁶ and SO₂R⁶,and wherein said aryl and heteroaryl rings are optionally furthersubstituted with one or more groups independently selected from F, Cl,CF₃, CF₃O—, HCF₂O—, alkyl, heteroalkyl and ArO—;

R¹, R³ and R⁴ are independently selected from H, alkyl, alkenyl,alkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, aryland heteroaryl, wherein said alkyl, alkenyl, alkynyl, heteroalkyl,cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl and heteroaryl areoptionally substituted with one or more groups independently selectedfrom alkyl, alkenyl, alkynyl, F, Cl, Br, I, CN, OR⁶, NR⁶R⁷, C(═O)R⁶,C(═O)OR⁶, OC(═O)R⁶, C(═O)NR⁶R⁷, (C₁-C₆ alkyl)amino, CH₃OCH₂O—,R⁶OC(═O)CH═CH₂—, NR⁶SO₂R⁷, SR⁶ and SO₂R⁶,

or R³ and R⁴ together with the atom to which they are attached form asaturated or partially unsaturated carbocyclic ring, wherein saidcarbocyclic ring is optionally substituted with one or more groupsindependently selected from alkyl, alkenyl, alkynyl, F, Cl, Br, I, CN,OR⁶, NR⁶R⁷, C(═O)R⁶, C(═O)OR⁶, OC(═O)R⁶, C(═O)NR⁶R⁷, (C₁-C₆ alkyl)amino,CH₃OCH₂O—, R⁶OC(═O)CH═CH₂—, NR⁶SO₂R⁷, SR⁶ and SO₂R⁶;

R² and R⁸ are independently selected from H, OR⁶, NR⁶R⁷, alkyl, alkenyl,alkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, aryland heteroaryl, wherein said alkyl, alkenyl, alkynyl, heteroalkyl,cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl and heteroaryl areoptionally substituted with one or more groups independently selectedfrom alkyl, alkenyl, alkynyl, F, Cl, Br, I, CN, OR⁶, NR⁶R⁷, C(═O)R⁶,C(═O)OR⁶, OC(═O)R⁶, C(═O)NR⁶R⁷, (C₁-C₆ alkyl)amino, CH₃OCH₂O—,R⁶OC(═O)CH═CH₂—, NR⁶SO₂R⁷, SR⁶ and SO₂R⁶;

R^(5a), R^(5b), and R^(5c) are independently H, F, Cl, Br, I, OMe, CH₃,CH₂F, CHF₂ or CF₃; and

R⁶ and R⁷ are independently selected from H, alkyl, alkenyl, alkynyl,heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl andheteroaryl, wherein said alkyl, alkenyl, alkynyl, heteroalkyl,cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl and heteroaryl areoptionally substituted with one or more groups independently selectedfrom alkyl, alkenyl, alkynyl, F, Cl, Br, I, CN, OR⁶, NR⁶R⁷, C(═O)R⁶,C(═O)OR⁶, OC(═O)R⁶, C(═O)NR⁶R⁷, (C₁-C₆ alkyl)amino, CH₃OCH₂O—,R⁶OC(═O)CH═CH₂—, NR⁶SO₂R⁷, SR⁶ and SO₂R⁶,

or R⁶ and R⁷ together with the atom to which they are attached form asaturated or partially unsaturated heterocyclic ring, wherein saidheterocyclic ring is optionally substituted with one or more groupsindependently selected from alkyl, alkenyl, alkynyl, F, Cl, Br, I, CN,OR⁶, NR⁶R⁷, C(═O)R⁶, C(═O)OR⁶, OC(═O)R⁶, C(═O)NR⁶R⁷, (C₁-C₆ alkyl)amino,CH₃OCH₂O—, R⁶OC(═O)CH═CH₂—, NR⁶SO₂R⁷, SR⁶ and SO₂R⁶.

In certain embodiments, R¹, R³ and R⁴ are each hydrogen.

In certain embodiments, R^(5a), R^(5b) and R^(5c) are each hydrogen.

In certain embodiments of said compound of Formula I, R² is OR⁶. Incertain embodiments, R⁶ is alkyl, such as (1-4C)alkyl. In particularembodiments, R⁶ is ethyl.

In certain embodiments of said compound of Formula I, R² is NR⁶R⁷. Incertain embodiments, R⁶ and R⁷ are independently H, alkyl, such as(1-6C)alkyl, or heteroalkyl, such as (1-4C)alkoxy(2-4C)alkyl. Inparticular embodiments, R⁶ and R⁷ are independently H, ethyl, propyl, orCH₂CH₂OCH₃.

In certain embodiments of said compound of Formula I, Y is aryl, such asphenyl. In certain embodiments, said aryl is substituted with C(═O)R⁸,such as in para-R⁸C(═O)phenyl. In certain embodiments, R⁸ is OR⁶, NR⁶R⁷or heterocycloalkyl. In certain embodiments, R⁶ and R⁷ are independentlyH or alkyl, such as (1-6C)alkyl. In certain other embodiments, R⁶ and R⁷together with the nitrogen atom to which they are attached form a 4-6membered azacycloalkyl ring, such as pyrrolidinyl. In particularembodiments, Y is

In certain embodiments of said compound of Formula I, Y is CF₂CF₃.

The term “alkyl” as used herein refers to a saturated linear orbranched-chain monovalent hydrocarbon radical having one to twelve,including one to ten carbon atoms, one to six carbon atoms and one tofour carbon atoms, wherein the alkyl radical may be optionallysubstituted independently with one or more substituents described below.Examples of alkyl radicals include C₁-C₁₂ hydrocarbon moieties such as,but not limited to: methyl (Me, —CH₃), ethyl (Et, —CH₂CH₃), 1-propyl(n-Pr, n-propyl, —CH₂CH₂CH₃), 2-propyl (i-Pr, i-propyl, —CH(CH₃)₂),1-butyl (n-Bu, n-butyl, —CH₂CH₂CH₂CH₃), 2-methyl-1-propyl (1-Bu,i-butyl, —CH₂CH(CH₃)₂), 2-butyl (s-Bu, s-butyl, —CH(CH₃)CH₂CH₃),2-methyl-2-propyl (t-Bu, t-butyl, —C(CH₃)₃), 1-pentyl (n-pentyl,—CH₂CH₂CH₂CH₂CH₃), 2-pentyl (—CH(CH₃) CH₂CH₂CH₃), 3-pentyl(—CH(CH₂CH₃)₂), 2-methyl-2-butyl (—C(CH₃)₂CH₂CH₃), 3-methyl-2-butyl(—CH(CH₃)CH(CH₃)₂), 3-methyl-1-butyl (—CH₂CH₂CH(CH₃)₂), 2-methyl-1-butyl(—CH₂CH(CH₃)CH₂CH₃), 1-hexyl (—CH₂CH₂CH₂CH₂CH₂CH₃), 2-hexyl(—CH(CH₃)CH₂CH₂ CH₂CH₃), 3-hexyl (—CH(CH₂CH₃)(CH₂CH₂CH₃)),2-methyl-2-pentyl (—C(CH₃)₂CH₂CH₂ CH₃), 3-methyl-2-pentyl(—CH(CH₃)CH(CH₃)CH₂CH₃), 4-methyl-2-pentyl (—CH(CH₃)CH₂ CH(CH₃)₂),3-methyl-3-pentyl (—C(CH₃)(CH₂CH₃)₂), 2-methyl-3-pentyl (—CH(CH₂CH₃)CH(CH₃)₂), 2,3-dimethyl-2-butyl (—C(CH₃)₂CH(CH₃)₂), 3,3-dimethyl-2-butyl(—CH(CH₃)C (CH₃)₃, 1-heptyl, and 1-octyl.

The term “alkenyl” refers to a linear or branched-chain monovalenthydrocarbon radical having two to 10 carbon atoms, including two to sixcarbon atoms and two to four carbon atoms, and at least one double bond,and includes, but is not limited to, ethenyl, propenyl, 1-but-3-enyl,1-pent-3-enyl, 1-hex-5-enyl and the like, wherein the alkenyl radicalmay be optionally substituted independently with one or moresubstituents described herein, and includes radicals having “cis” and“trans” orientations, or alternatively, “E” and “Z” orientations. Theterm “alkenyl” includes allyl.

The term “alkynyl” refers to a linear or branched monovalent hydrocarbonradical of two to twelve carbon atoms, including two to 10 carbon atoms,two to six carbon atoms and two to four carbon atoms, containing atleast one triple bond. Examples include, but are not limited to,ethynyl, propynyl, butynyl, pentyn-2-yl and the like, wherein thealkynyl radical may be optionally substituted independently with one ormore substituents described herein.

The terms “carbocycle,” “carbocyclyl,” or “cycloalkyl” are usedinterchangeably herein and refer to saturated or partially unsaturatedcyclic hydrocarbon radical having from three to twelve carbon atoms,including from three to ten carbon atoms and from three to six carbonatoms. The term “cycloalkyl” includes monocyclic and polycyclic (e.g.,bicyclic and tricyclic) cycloalkyl structures, wherein the polycyclicstructures optionally include a saturated or partially unsaturatedcycloalkyl fused to a saturated or partially unsaturated cycloalkyl orheterocycloalkyl ring or an aryl or heteroaryl ring. Examples ofcycloalkyl groups include, but are not limited to, cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and the like. Bicycliccarbocycles have 7 to 12 ring atoms, e.g. arranged as a bicyclo[4,5],[5,5], [5,6] or [6,6] system, or 9 or 10 ring atoms arranged as abicyclo[5,6] or [6,6] system, or as bridged systems such asbicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, and bicyclo[3.2.2]nonane.The cycloalkyl may be optionally substituted independently at one ormore substitutable positions with one or more substituents describedherein. Such cycloalkyl groups may be optionally substituted with, forexample, one or more groups independently selected from C₁-C₆ alkyl,C₁-C₆ alkoxy, halogen, hydroxy, cyano, nitro, amino, mono(C₁-C₆)alkylamino, di(C₁-C₆)alkylamino, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₁-C₆haloalkyl, C₁-C₆ haloalkoxy, amino(C₁-C₆)alkyl,mono(C₁-C₆)alkylamino(C₁-C₆)alkyl and di(C₁-C₆)alkylamino(C₁-C₆)alkyl.

The term “cycloalkenyl” refers to a partially unsaturated cyclichydrocarbon radical having from three to ten carbon atoms, includingfrom three to six carbon atoms, and having at least one double bondwithin the carbocycle.

The term “heteroalkyl” refers to saturated linear or branched-chainmonovalent hydrocarbon radical of one to twelve carbon atoms, includingfrom one to six carbon atoms and from one to four carbon atoms, whereinat least one of the carbon atoms is replaced with a heteroatom selectedfrom N, O, or S, and wherein the radical may be a carbon radical orheteroatom radical (i.e., the heteroatom may appear in the middle or atthe end of the radical). The heteroalkyl radical may be optionallysubstituted independently with one or more substituents describedherein. The term “heteroalkyl” encompasses alkoxy and heteroalkoxyradicals.

The terms “heterocycloalkyl,” “heterocycle” and “heterocyclyl” are usedinterchangeably herein and refer to a saturated or partially unsaturatedcarbocyclic radical of 3 to 8 ring atoms in which at least one ring atomis a heteroatom selected from nitrogen, oxygen and sulfur, the remainingring atoms being C, where one or more ring atoms may be optionallysubstituted independently with one or more substituents described below.The radical may be a carbon radical or heteroatom radical. The term“heterocycle” includes heterocycloalkoxy. The term further includesfused ring systems which include a heterocycle fused to an aromaticgroup. “Heterocycloalkyl” also includes radicals where heterocycleradicals are fused with aromatic or heteroaromatic rings. Examples ofheterocycloalkyl rings include, but are not limited to, pyrrolidinyl,tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, tetrahydropyranyl,dihydropyranyl, tetrahydrothiopyranyl, piperidino, morpholino,thiomorpholino, thioxanyl, piperazinyl, homopiperazinyl, azetidinyl,oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl,diazepinyl, thiazepinyl, 1,2,3,6-tetrahydropyridinyl, 2-pyrrolinyl,3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl,1,3-dioxolanyl, pyrazolinyl, dithianyl, dithiolanyl, dihydropyranyl,dihydrothienyl, dihydrofuranyl, pyrazolidinylimidazolinyl,imidazolidinyl, 3-azabicyco[3.1.0]hexanyl, 3-azabicyclo[4.1.0]heptanyl,azabicyclo[2.2.2]hexanyl, 3H-indolyl quinolizinyl and N-pyridyl ureas.Spiro moieties are also included within the scope of this definition.The foregoing groups, as derived from the groups listed above, may beC-attached or N-attached where such is possible. For instance, a groupderived from pyrrole may be pyrrol-1-yl (N-attached) or pyrrol-3-yl(C-attached). Further, a group derived from imidazole may beimidazol-1-yl (N-attached) or imidazol-3-yl (C-attached). An example ofa heterocyclic group wherein 2 ring carbon atoms are substituted withoxo (═O) moieties is 1,1-dioxo-thiomorpholinyl. The heterocycle groupsherein are unsubstituted or, as specified, substituted in one or moresubstitutable positions with various groups. For example, suchheterocycle groups may be optionally substituted with, for example, oneor more groups independently selected from C₁-C₆ alkyl, C₁-C₆ alkoxy,halogen, hydroxy, cyano, nitro, amino, mono(C₁-C₆)alkylamino,di(C₁-C₆)alkylamino, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₁-C₆ haloalkyl,C₁-C₆ haloalkoxy, amino(C₁-C₆)alkyl, mono(C₁-C₆)alkylamino(C₁-C₆)alkylor di(C₁-C₆)alkylamino(C₁-C₆)alkyl.

The term “aryl” refers to a monovalent aromatic carbocyclic radicalhaving a single ring (e.g., phenyl), multiple rings (e.g., biphenyl), ormultiple condensed rings in which at least one is aromatic, (e.g.,1,2,3,4-tetrahydronaphthyl, naphthyl, etc.), which is optionallysubstituted with one or more substituents independently selected from,for example, halogen, lower alkyl, lower alkoxy, trifluoromethyl, aryl,heteroaryl and hydroxy.

The term “heteroaryl” refers to a monovalent aromatic radical of 5-, 6-,or 7-membered rings and includes fused ring systems (at least one ofwhich is aromatic) of 5-10 atoms containing at least one and up to fourheteroatoms selected from nitrogen, oxygen, and sulfur. Examples ofheteroaryl groups are pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl,triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl,oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, indolyl,benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl,phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl,oxadiazolyl, triazolyl, thiadiazolyl, thiadiazolyl, furazanyl,benzofurazanyl, benzothiophenyl, benzothiazolyl, benzoxazolyl,quinazolinyl, quinoxalinyl, naphthyridinyl, and furopyridinyl. Spiromoieties are also included within the scope of this definition.Heteroaryl groups are optionally substituted with one or moresubstituents independently selected from, for example, halogen, loweralkyl, lower alkoxy, haloalkyl, aryl, heteroaryl, and hydroxy.

The term “halogen” represents fluorine, bromine, chlorine, and iodine.

The term “oxo” represents ═O.

In general, the various moieties or functional groups of the compoundsof Formula I may be optionally substituted by one or more substituents.Examples of substituents suitable for purposes of this inventioninclude, but are not limited to, oxo, halogen, cyano, nitro,trifluoromethyl, difluoromethoxy, trifluoromethoxy, azido, —NR″SO₂R′,—SO₂NR′R″, —C(O)R′, —C(O)OR′, —OC(O)R′, —NR″C(O)OR′, —NR″C(O)R′,—C(O)NR′R″, —NR′R″, —NR″′C(O)N′R″, —NR″′C(NCN)NR′R″, —OR′, aryl,heteroaryl, arylalkyl, heteroarylalkyl, heterocyclyl, andheterocyclylalkyl, where R′, R″ and R″′ are independently H, alkyl,heteroalkyl, cycloalkyl, heterocycloalkyl, alkenyl, alkynyl, aryl orheteroaryl.

It is to be understood that in instances where two or more radicals areused in succession to define a substituent attached to a structure, thefirst named radical is considered to be terminal and the last namedradical is considered to be attached to the structure in question. Thus,for example, an arylalkyl radical is attached to the structure inquestion by the alkyl group.

The compounds of this invention may possess one or more asymmetriccenters; such compounds can therefore be produced as individual (R)- or(S)-stereoisomers or as mixtures thereof. Unless indicated otherwise,the description or naming of a particular compound in the specificationand claims is intended to include both individual enantiomers,diastereomers mixtures, racemic or otherwise, thereof. Accordingly, thisinvention also includes all such isomers, including diastereomericmixtures, pure diastereomers and pure enantiomers of the Formula I.Diastereomeric mixtures can be separated into their individualdiastereomers on the basis of their physical chemical differences bymethods known to those skilled in the art, for example, bychromatography or fractional crystallization. Enantiomers can beseparated by converting the enantiomer mixture into a diastereomericmixture by reaction with an appropriate optically active compound (e.g.,alcohol), separating the diastereomers and converting (e.g.,hydrolyzing) the individual diastereomers to the corresponding pureenantiomers. Enantiomers can also be separated by use of a chiral HPLCcolumn. Methods for the determination of stereochemistry and theseparation of stereoisomers are well known in the art (see discussion inChapter 4 of “Advanced Organic Chemistry”, 4th edition, J. March, JohnWiley and Sons, New York, 1992).

In the structures shown herein, where the stereochemistry of anyparticular chiral atom is not specified, then all stereoisomers arecontemplated and included as the compounds of the invention. Wherestereochemistry is specified by a solid wedge or dashed linerepresenting a particular configuration, then that stereoisomer is sospecified and defined.

A single stereoisomer, e.g. an enantiomer, substantially free of itsstereoisomer may be obtained by resolution of the racemic mixture usinga method such as formation of diastereomers using optically activeresolving agents (Eliel, E. and Wilen, S. Stereochemistry of OrganicCompounds, John Wiley & Sons, Inc., New York, 1994; Lochmuller, C. H.,(1975) J. Chromatogr., 113(3):283-302). Racemic mixtures of chiralcompounds of the invention can be separated and isolated by any suitablemethod, including: (1) formation of ionic, diastereomeric salts withchiral compounds and separation by fractional crystallization or othermethods, (2) formation of diastereomeric compounds with chiralderivatizing reagents, separation of the diastereomers, and conversionto the pure stereoisomers, and (3) separation of the substantially pureor enriched stereoisomers directly under chiral conditions. See: DrugStereochemistry, Analytical Methods and Pharmacology, Irving W. Wainer,Ed., Marcel Dekker, Inc., New York (1993).

Under method (1), diastereomeric salts can be formed by reaction ofenantiomerically pure chiral bases such as brucine, quinine, ephedrine,strychnine, α-methyl-β-phenylethylamine (amphetamine), and the like withasymmetric compounds bearing acidic functionality, such as carboxylicacid and sulfonic acid. The diastereomeric salts may be induced toseparate by fractional crystallization or ionic chromatography. Forseparation of the optical isomers of amino compounds, addition of chiralcarboxylic or sulfonic acids, such as camphorsulfonic acid, tartaricacid, mandelic acid, or lactic acid can result in formation of thediastereomeric salts.

Alternatively, by method (2), the substrate to be resolved is reactedwith one enantiomer of a chiral compound to form a diastereomeric pair(E. and Wilen, S. “Stereochemistry of Organic Compounds”, John Wiley &Sons, Inc., 1994, p. 322). Diastereomeric compounds can be formed byreacting asymmetric compounds with enantiomerically pure chiralderivatizing reagents, such as menthyl derivatives, followed byseparation of the diastereomers and hydrolysis to yield the pure orenriched enantiomer. A method of determining optical purity involvesmaking chiral esters, for example a menthyl ester such as (−) menthylchloroformate, in the presence of base, or Mosher ester,α-methoxy-α-(trifluoromethyl)phenyl acetate (Jacob III, (1982) J. Org.Chem. 47:4165), of the racemic mixture, and analyzing the NMR spectrumfor the presence of the two atropisomeric enantiomers or diastereomers.Stable diastereomers of atropisomeric compounds can be separated andisolated by normal- and reverse-phase chromatography following methodsfor separation of atropisomeric naphthyl-isoquinolines (WO 96/15111). Bymethod (3), a racemic mixture of two enantiomers can be separated bychromatography using a chiral stationary phase (Chiral LiquidChromatography (1989) W. J. Lough, Ed., Chapman and Hall, New York;Okamoto, (1990) J. of Chromatogr. 513:375-378). Enriched or purifiedenantiomers can be distinguished by methods used to distinguish otherchiral molecules with asymmetric carbon atoms, such as optical rotationand circular dichroism.

In addition to compounds of the Formula I, the invention also includessolvates, pharmaceutically acceptable prodrugs, pharmaceutically activemetabolites, solvates, and pharmaceutically acceptable salts of suchcompounds.

The term “solvate” refers to an aggregate of a molecule with one or moresolvent molecules.

A “pharmaceutically acceptable prodrug” is a compound that may beconverted under physiological conditions or by solvolysis to thespecified compound or to a pharmaceutically acceptable salt of suchcompound. Prodrugs include compounds wherein an amino acid residue, or apolypeptide chain of two or more (e.g., two, three or four) amino acidresidues is covalently joined through an amide or ester bond to a freeamino, hydroxy or carboxylic acid group of compounds of the presentinvention. The amino acid residues include but are not limited to the 20naturally occurring amino acids commonly designated by three lettersymbols and also includes phosphoserine, phosphothreonine,phosphotyrosine, 4-hydroxyproline, hydroxylysine, demosine, isodemosine,gamma-carboxyglutamate, hippuric acid, octahydroindole-2-carboxylicacid, statine, 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid,penicillamine, ornithine, 3-methylhistidine, norvaline, beta-alanine,gamma-aminobutyric acid, cirtulline, homocysteine, homoserine,methyl-alanine, para-benzoylphenylalanine, phenylglycine,propargylglycine, sarcosine, methionine sulfone and tert-butylglycine.Particular examples of prodrugs of this invention include a compound ofFormula I covalently joined to a phosphate residue or a valine residue.

Additional types of prodrugs are also encompassed. For instance, freecarboxyl groups can be derivatized as amides or alkyl esters. As anotherexample, compounds of this invention comprising free hydroxy groups maybe derivatized as prodrugs by converting the hydroxy group into groupssuch as, but not limited to, phosphate ester, hemisuccinate,dimethylaminoacetate, or phosphoryloxymethyloxycarbonyl groups, asoutlined in Advanced Drug Delivery Reviews, (1996) 19:115. Carbamateprodrugs of hydroxy and amino groups are also included, as are carbonateprodrugs, sulfonate esters and sulfate esters of hydroxy groups.Derivatization of hydroxy groups as (acyloxy)methyl and (acyloxy)ethylethers wherein the acyl group may be an alkyl ester, optionallysubstituted with groups including, but not limited to, ether, amine andcarboxylic acid functionalities, or where the acyl group is an aminoacid ester as described above, are also encompassed. Prodrugs of thistype are described in J. Med. Chem., (1996) 39:10. More specificexamples include replacement of the hydrogen atom of the alcohol groupwith a group such as (C₁-C₆)alkanoyloxymethyl,1-((C₁-C₆)alkanoyloxy)ethyl, 1-methyl-1-((C₁-C₆)alkanoyloxy)ethyl,(C₁-C₆)alkoxycarbonyl oxymethyl, N—(C₁-C₆)alkoxycarbonylaminomethyl,succinoyl, (C₁-C₆)alkanoyl, α-amino (C₁-C₄)alkanoyl, arylacyl andα-aminoacyl, or α-aminoacyl-α-aminoacyl, where each α-aminoacyl group isindependently selected from the naturally occurring L-amino acids,P(O)(OH)₂, —P(O)(O(C₁-C₆)alkyl)₂ or glycosyl (the radical resulting fromthe removal of a hydroxyl group of the hemiacetal form of acarbohydrate).

Free amines can also be derivatized as amides, sulfonamides orphosphonamides. All of these prodrug moieties may incorporate groupsincluding, but not limited to, ether, amine and carboxylic acidfunctionalities. For example, a prodrug can be formed by the replacementof a hydrogen atom in the amine group with a group such as R-carbonyl,RO-carbonyl, NRR′-carbonyl where R and R′ are each independently(C₁-C₁₀)alkyl, (C₃-C₇)cycloalkyl, benzyl, or R-carbonyl is a naturalα-aminoacyl or natural α-aminoacyl-natural α-aminoacyl, —C(OH)C(O)OYwherein Y is H, (C₁-C₆)alkyl or benzyl, —C(OY₀)Y₁ wherein Y₀ is(C₁-C₄)alkyl and Y₁ is (C₁-C₆)alkyl, carboxy(C₁-C₆)alkyl,amino(C₁-C₄)alkyl or mono-N— or di-N,N—(C₁-C₆)alkylaminoalkyl, —C(Y₂)Y₃wherein Y₂ is H or methyl and Y₃ is mono-N— or di-N,N—(C₁-C₆)alkylamino,morpholino, piperidin-1-yl or pyrrolidin-1-yl.

For additional examples of prodrug derivatives, see, for example, a)Design of Prodrugs, edited by H. Bundgaard, (Elsevier, 1985) and Methodsin Enzymology, Vol. 42, p. 309-396, edited by K. Widder, et al.(Academic Press, 1985); b) A Textbook of Drug Design and Development,edited by Krogsgaard-Larsen and H. Bundgaard, Chapter 5 “Design andApplication of Prodrugs,” by H. Bundgaard p. 113-191 (1991); c) H.Bundgaard, Advanced Drug Delivery Reviews, (1992); 8:1-38 d) H.Bundgaard, et al., Journal of Pharmaceutical Sciences, (1988) 77:285;and e) N. Kakeya, et al., Chem. Pharm. Bull., (1984) 32:692, each ofwhich is specifically incorporated herein by reference.

A “pharmaceutically active metabolite” is a pharmacologically activeproduct produced through metabolism in the body of a specified compoundor salt thereof. Metabolites of a compound may be identified usingroutine techniques known in the art and their activities determinedusing tests such as those described herein.

Prodrugs and active metabolites of a compound may be identified usingroutine techniques known in the art.

A “pharmaceutically acceptable salt,” unless otherwise indicated,includes salts that retain the biological effectiveness of the freeacids and bases of the specified compound and that are not biologicallyor otherwise undesirable. A compound of the invention may possess asufficiently acidic, a sufficiently basic, or both functional groups,and accordingly react with any of a number of inorganic or organicbases, and inorganic and organic acids, to form a pharmaceuticallyacceptable salt. Examples of pharmaceutically acceptable salts includethose salts prepared by reaction of the compounds of the presentinvention with a mineral or organic acid or an inorganic base, suchsalts including sulfates, pyrosulfates, bisulfates, sulfites,bisulfites, phosphates, monohydrogenphosphates, dihydrogenphosphates,metaphosphates, pyrophosphates, chlorides, bromides, iodides, acetates,propionates, decanoates, caprylates, acrylates, formates, isobutyrates,caproates, heptanoates, propiolates, oxalates, malonates, succinates,suberates, sebacates, fumarates, maleates, butyn-1,4-dioates,hexyne-1,6-dioates, benzoates, chlorobenzoates, methylbenzoates,dinitrobenzoates, hydroxybenzoates, methoxybenzoates, phthalates,sulfonates, xylenesulfonates, pheylacetates, phenylpropionates,phenylbutyrates, citrates, lactates, γ-hydroxybutyrates, glycollates,tartrates, methanesulfonates, propanesulfonates,naphthalene-1-sulfonates, naphthalene-2-sulfonates, and mandelates.Since a single compound of the present invention may include more thanone acidic or basic moieties, the compounds of the present invention mayinclude mono, di or tri-salts in a single compound.

If the inventive compound is a base, the desired pharmaceuticallyacceptable salt may be prepared by any suitable method available in theart, for example, treatment of the free base with an acidic compound,particularly an inorganic acid, such as hydrochloric acid, hydrobromicacid, sulfuric acid, nitric acid, phosphoric acid and the like, or withan organic acid, such as acetic acid, maleic acid, succinic acid,mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid,glycolic acid, salicylic acid, a pyranosidyl acid such as glucuronicacid or galacturonic acid, an alpha hydroxy acid such as citric acid ortartaric acid, an amino acid such as aspartic acid or glutamic acid, anaromatic acid such as benzoic acid or cinnamic acid, a sulfonic acidsuch as p-toluenesulfonic acid or ethanesulfonic acid, or the like.

If the inventive compound is an acid, the desired pharmaceuticallyacceptable salt may be prepared by any suitable method, for example,treatment of the free acid with an inorganic or organic base. Examplesof suitable inorganic salts include those formed with alkali andalkaline earth metals such as lithium, sodium, potassium, barium andcalcium. Examples of suitable organic base salts include, for example,ammonium, dibenzylammonium, benzylammonium, 2-hydroxyethylammonium,bis(2-hydroxyethyl) ammonium, phenylethylbenzylamine,dibenzylethylenediamine, and the like salts. Other salts of acidicmoieties may include, for example, those salts formed with procaine,quinine and N-methylglucosamine, plus salts formed with basic aminoacids such as glycine, ornithine, histidine, phenylglycine, lysine andarginine.

The present invention also provides salts of compounds of Formula Iwhich are not necessarily pharmaceutically acceptable salts, but whichmay be useful as intermediates for preparing and/or purifying compoundsof Formula I and/or for separating enantiomers of compounds of FormulaI.

The inventive compounds may be prepared using the reaction routes andsynthesis schemes as described in Scheme I, employing the techniquesavailable in the art using starting materials that are readilyavailable, or can be synthesized according to the methods described inthe Examples and FIGS. 1-4 or using methods known in the art.

In Scheme I, compounds of Formula II may be prepared from an alkyl areneof Formula I by treatment with dimethylformamide dimethyl acetal with orwithout the use of pyrollidine (J. Org. Chem., (1986), 51(26),5106-5110) in DMF at 70-90° C. The crude intermediate (not shown) may becleaved to the aldehyde of Formula II with NaIO₄ in THF/pH 7.2 phosphatebuffer at or around room temperature. The aldehyde of Formula II may beolefinated with phosphonium ylid in toluene at temperatures ranging from70 to 110° C. (1-16 hours) to give compounds of Formula III. Compoundsof Formula IV can be prepared from a compound of Formula III using ironpowder in acetic acid. The reaction may be conducted at temperaturesbetween around 90° C. for about 3-14 hours.

It is noted that some of the preparations of compounds of Formula Idescribed herein may require protection of remote functionalities. Theneed for such protection will vary depending on the nature of thefunctionality and the conditions used in the preparation methods and canbe readily determined by those skilled in the art. Suchprotection/deprotection methods are well known to those skilled in theart.

The compounds of the invention find use in a variety of applications.For example, in certain aspects the invention provides methods formodulating TLR7- and/or TLR8-mediated signaling. The methods of theinvention are useful, for example, when it is desirable to alter TLR7-and/or TLR8-mediated signaling in response to a suitable TLR7 and/orTLR8 ligand or a TLR7 and/or TLR8 signaling agonist.

As used herein, the terms “TLR7 and/or TLR8 ligand,” “ligand for TLR7and/or TLR8,” and “TLR7 and/or TLR8 signaling agonist” refer to amolecule, other than a compound of Formula I, that interacts directly orindirectly with TLR7 and/or TLR8 through a TLR7 and/or TLR8 domain otherthan a TIR8 domain, and induces TLR7- and/or TLR8-mediated signaling. Incertain embodiments, a TLR7 and/or TLR8 ligand is a natural ligand,i.e., a TLR7 and/or TLR8 ligand that is found in nature. In certainembodiments, a TLR7 and/or TLR8 ligand refers to a molecule other than anatural ligand of TLR7 and/or TLR8, e.g., a molecule prepared by humanactivity.

The term “modulate” as used herein with respect to the TLR7 and/or TLR8receptors means the mediation of a pharmacodynamic response in a subjectby (i) inhibiting or activating the receptor, or (ii) directly orindirectly affecting the normal regulation of the receptor activity.Compounds that modulate the receptor activity include agonists,antagonists, mixed agonists/antagonists and compounds that directly orindirectly affect regulation of the receptor activity.

The term “agonist” refers to a compound that, in combination with areceptor (e.g., a TLR), can produce a cellular response. An agonist maybe a ligand that directly binds to the receptor. Alternatively, anagonist may combine with a receptor indirectly by, for example, (a)forming a complex with another molecule that directly binds to thereceptor, or (b) otherwise resulting in the modification of anothercompound so that the other compound directly binds to the receptor. Anagonist may be referred to as an agonist of a particular TLR (e.g., aTLR7 and/or TLR8 agonist).

The term “antagonist” as used herein refers to a compound that competeswith an agonist or inverse agonist for binding to a receptor, therebyblocking the action of an agonist or inverse agonist on the receptor.However, an antagonist has no effect on constitutive receptor activity.More specifically, an antagonist is a compound that inhibits theactivity of TRL7 or TLR8 at the TLR7 or TLR8 receptor, respectively.

“Inhibit” refers to any measurable reduction of biological activity.Thus, as used herein, “inhibit” or “inhibition” may be referred to as apercentage of a normal level of activity.

In one aspect of this invention, a method of treating a condition ordisorder treatable by modulation of TLR7- and/or TLR8-mediated cellularactivities in a subject comprises administering to said subject acomposition of Formula I in an amount effective to treat the conditionor disorder. The term “TLR7- and/or TLR8-mediated” refers to abiological or biochemical activity that results from TLR7- and/or TLR8function.

Conditions and disorders that can be treated by the methods of thisinvention include, but are not limited to, cancer, immunecomplex-associated diseases, inflammatory disorders, immunodeficiency,graft rejection, graft-versus-host disease, allergies, asthma,infection, and sepsis. More specifically, methods useful in thetreatment of conditions involving autoimmunity, inflammation, allergy,asthma, graft rejection, and GvHD generally will employ compounds ofFormula I that inhibit TLR7- and/or TLR8-mediated signaling in responseto a suitable TLR7 and/or TLR8 ligand or signaling agonist.Alternatively, methods useful in the treatment of conditions involvinginfection, cancer, and immunodeficiency generally will employ compoundsof Formula I that augment TLR7- and/or TLR8-mediated signaling inresponse to a suitable TLR7 and/or TLR8 ligand. In some instances thecompositions can be used to inhibit or promote TLR7- and/orTLR8-mediated signaling in response to a TLR7 and/or TLR8 ligand orsignaling agonist. In other instances the compositions can be used toinhibit or promote TLR7- and/or TLR8-mediated immunostimulation in asubject.

The term “treating” as used herein, unless otherwise indicated, means atleast the mitigation of a disease condition in a mammal, such as ahuman, and includes, but is not limited to, modulating and/or inhibitingthe disease condition, and/or alleviating the disease condition to whichsuch term applies, or one or more symptoms of such disorder orcondition. The term “treatment,” as used herein, unless otherwiseindicated, refers to the act of treating as “treating” is definedimmediately above.

As used herein, the terms “autoimmune disease,” “autoimmune disorder”and “autoimmunity” refer to immunologically mediated acute or chronicinjury to a tissue or organ derived from the host. The terms encompassboth cellular and antibody-mediated autoimmune phenomena, as well asorgan-specific and organ-nonspecific autoimmunity. Autoimmune diseasesinclude insulin-dependent diabetes mellitus, rheumatoid arthritis,systemic lupus erythematosus, multiple sclerosis, atherosclerosis, andinflammatory bowel disease. Autoimmune diseases also include, withoutlimitation, ankylosing spondylitis, autoimmune hemolytic anemia,Behget's syndrome, Goodpasture's syndrome, Graves' disease,Guillain-Barre syndrome, Hashimoto's thyroiditis, idiopathicthrombocytopenia, myasthenia gravis, pernicious anemia, polyarteritisnodosa, polymyositis/dermatomyositis, primary biliary sclerosis,psoriasis, sarcoidosis, sclerosing cholangitis, Sjogren's syndrome,systemic sclerosis (scleroderma and CREST syndrome), Takayasu'sarteritis, temporal arteritis, and Wegener's granulomatosis. Autoimmunediseases also include certain immune complex-associated diseases.

As used herein, the terms “cancer” and, “tumor” refer to a condition inwhich abnormally replicating cells of host origin are present in adetectable amount in a subject. The cancer can be a malignant ornon-malignant cancer. Cancers or tumors include, but are not limited to,biliary tract cancer; brain cancer; breast cancer; cervical cancer;choriocarcinoma; colon cancer; endometrial cancer; esophageal cancer;gastric (stomach) cancer; intraepithelial neoplasms; leukemias;lymphomas; liver cancer; lung cancer (e.g., small cell and non-smallcell); melanoma; neuroblastomas; oral cancer; ovarian cancer; pancreaticcancer; prostate cancer; rectal cancer; renal (kidney) cancer; sarcomas;skin cancer; testicular cancer; thyroid cancer; as well as othercarcinomas and sarcomas. Cancers can be primary or metastatic.

As used herein, the term “immune complex-associated disease” refers toany disease characterized by the production and/or tissue deposition ofimmune complexes (i.e., any conjugate including an antibody and anantigen specifically bound by the antibody), including, but not limitedto systemic lupus erythematosus (SLE) and related connective tissuediseases, rheumatoid arthritis, hepatitis C- and hepatitis B-relatedimmune complex disease (e.g., cryoglobulinemia), Behget's syndrome,autoimmune glomerulonephritides, and vasculopathy associated with thepresence of LDL/anti-LDL immune complexes.

As used herein, “immunodeficiency” refers to a disease or disorder inwhich the subject's immune system is not functioning in normal capacityor in which it would be useful to boost a subject's immune response, forexample to eliminate a tumor or cancer (e.g., tumors of the brain, lung(e.g., small cell and non-small cell), ovary, breast, prostate, colon,as well as other carcinomas and sarcomas) or an infection in a subject.The immunodeficiency can be acquired or it can be congenital.

As used herein, “graft rejection” refers to immunologically mediatedhyperacute, acute, or chronic injury to a tissue or organ derived from asource other than the host. The term thus encompasses both cellular andantibody-mediated rejection, as well as rejection of both allografts andxenografts.

“Graft-versus-host disease” (GvHD) is a reaction of donated bone marrowagainst a patient's own tissue. GVHD is seen most often in cases wherethe blood marrow donor is unrelated to the patient or when the donor isrelated to the patient but not a perfect match. There are two forms ofGVHD: an early form called acute GVHD that occurs soon after thetransplant when the white cells are on the rise and a late form calledchronic GVHD.

T_(H2)-mediated, atopic diseases include, but are not limited to, atopicdermatitis or eczema, eosinophilia, asthma, allergy, allergic rhinitis,and Ommen's syndrome.

As used herein, “allergy” refers to acquired hypersensitivity to asubstance (allergen). Allergic conditions include eczema, allergicrhinitis or coryza, hay fever, asthma, urticaria (hives) and foodallergies, and other atopic conditions.

As used herein, “asthma” refers to a disorder of the respiratory systemcharacterized by inflammation, narrowing of the airways and increasedreactivity of the airways to inhaled agents. Asthma is frequently,although not exclusively associated with atopic or allergic symptoms.For example, asthma can be precipitated by exposure to an allergen,exposure to cold air, respiratory infection, and exertion.

As used herein, the terms “infection” and, equivalently, “infectiousdisease” refer to a condition in which an infectious organism or agentis present in a detectable amount in the blood or in a normally steriletissue or normally sterile compartment of a subject. Infectiousorganisms and agents include viruses, bacteria, fungi, and parasites.The terms encompass both acute and chronic infections, as well assepsis.

As used herein, the term “sepsis” refers to the presence of bacteria(bacteremia) or other infectious organisms or their toxins in the blood(septicemia) or in other tissue of the body.

Further provided is a compound of Formula I, or a metabolite, solvate,tautomer, or pharmaceutically acceptable prodrug or salt thereof, foruse as a medicament in the treatment of the diseases or conditionsdescribed above in a mammal, for example, a human, suffering from suchdisease or condition. Also provided is the use of a compound of FormulaI, or a metabolite, solvate, tautomer, or pharmaceutically acceptableprodrug or salt thereof, in the preparation of a medicament for thetreatment of the diseases and conditions described above in a mammal,for example a human, suffering from such disorder.

This invention also encompasses pharmaceutical compositions containing acompound of Formula I and methods of treating conditions and disorderstreatable by modulation of TLR7- and/or TLR8-mediated cellularactivities by administering a pharmaceutical composition comprising acompound of Formula I, or a metabolite, solvate, tautomer, orpharmaceutically acceptable prodrug or salt thereof, to a patient inneed thereof.

In order to use a compound of Formula I or a solvate, metabolite,tautomer or pharmaceutically acceptable prodrug or salt thereof for thetherapeutic treatment (including prophylactic treatment) of mammalsincluding humans, it is normally formulated in accordance with standardpharmaceutical practice as a pharmaceutical composition. According tothis aspect of the invention there is provided a pharmaceuticalcomposition that comprises a compound of Formula I, or a solvate,metabolite, tautomer or pharmaceutically acceptable prodrug or saltthereof, as defined hereinbefore in association with a pharmaceuticallyacceptable diluent or carrier.

To prepare the pharmaceutical compositions according to this invention,a therapeutically or prophylactically effective amount of a compound ofFormula I or a solvate, metabolite tautomer or pharmaceuticallyacceptable prodrug or salt thereof (alone or together with an additionaltherapeutic agent as disclosed herein) is intimately admixed, forexample, with a pharmaceutically acceptable carrier according toconventional pharmaceutical compounding techniques to produce a dose. Acarrier may take a wide variety of forms depending on the form ofpreparation desired for administration, e.g., oral or parenteral.Examples of suitable carriers include any and all solvents, dispersionmedia, adjuvants, coatings, antibacterial and antifungal agents,isotonic and absorption delaying agents, sweeteners, stabilizers (topromote long term storage), emulsifiers, binding agents, thickeningagents, salts, preservatives, solvents, dispersion media, coatings,antibacterial and antifungal agents, isotonic and absorption delayingagents, flavoring agents, and miscellaneous materials such as buffersand absorbents that may be needed in order to prepare a particulartherapeutic composition. The use of such media and agents withpharmaceutically active substances is well known in the art. Exceptinsofar as any conventional media or agent is incompatible with acompound of Formula I, its use in the therapeutic compositions andpreparations is contemplated. Supplementary active ingredients can alsobe incorporated into the compositions and preparations as describedherein.

The compositions of the invention may be in a form suitable for oral use(for example as tablets, lozenges, hard or soft capsules, aqueous oroily suspensions, emulsions, dispersible powders or granules, syrups orelixirs), for topical use (for example as creams, ointments, gels, oraqueous or oily solutions or suspensions), for administration byinhalation (for example as a finely divided powder or a liquid aerosol),for administration by insufflation (for example as a finely dividedpowder) or for parenteral administration (for example as a sterileaqueous or oily solution for intravenous, subcutaneous, or intramusculardosing or as a suppository for rectal dosing). For example, compositionsintended for oral use may contain, for example, one or more coloring,sweetening, flavoring and/or preservative agents.

Suitable pharmaceutically-acceptable excipients for a tablet formulationinclude, for example, inert diluents such as lactose, sodium carbonate,calcium phosphate or calcium carbonate, granulating and disintegratingagents such as corn starch or algenic acid; binding agents such asstarch; lubricating agents such as magnesium stearate, stearic acid ortalc; preservative agents such as ethyl or propyl p-hydroxybenzoate, andanti-oxidants, such as ascorbic acid. Tablet formulations may beuncoated or coated either to modify their disintegration and thesubsequent absorption of the active ingredient within thegastrointestinal tract, or to improve their stability and/or appearance,in either case, using conventional coating agents and procedures wellknown in the art.

Compositions for oral use may be in the form of hard gelatin capsules inwhich the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules in which the active ingredient is mixed with water oran oil such as peanut oil, liquid paraffin, or olive oil.

Aqueous suspensions generally contain the active ingredient in finelypowdered form together with one or more suspending agents, such assodium carboxymethylcellulose, methylcellulose,hydroxypropylmethylcellulose, sodium alginate, polyvinyl-pyrrolidone,gum tragacanth and gum acacia; dispersing or wetting agents such aslecithin or condensation products of an alkylene oxide with fatty acids(for example polyoxethylene stearate), or condensation products ofethylene oxide with long chain aliphatic alcohols, for exampleheptadecaethyleneoxycetanol, or condensation products of ethylene oxidewith partial esters derived from fatty acids and a hexitol such aspolyoxyethylene sorbitol monooleate, or condensation products ofethylene oxide with partial esters derived from fatty acids and hexitolanhydrides, for example polyethylene sorbitan monooleate. The aqueoussuspensions may also contain one or more preservatives (such as ethyl orpropyl p-hydroxybenzoate, anti-oxidants (such as ascorbic acid),coloring agents, flavoring agents, and/or sweetening agents (such assucrose, saccharine or aspartame).

Oily suspensions may be formulated by suspending the active ingredientin a vegetable oil (such as arachis oil, olive oil, sesame oil orcoconut oil) or in a mineral oil (such as liquid paraffin). The oilysuspensions may also contain a thickening agent such as beeswax, hardparaffin or cetyl alcohol. Sweetening agents such as those set outabove, and flavoring agents may be added to provide a palatable oralpreparation. These compositions may be preserved by the addition of ananti-oxidant such as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water generally contain the activeingredient together with a dispersing or wetting agent, suspending agentand one or more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.Additional excipients such as sweetening, flavoring and coloring agents,may also be present.

The pharmaceutical compositions of the invention may also be in the formof oil-in-water emulsions. The oily phase may be a vegetable oil, suchas olive oil or arachis oil, or a mineral oil, such as for exampleliquid paraffin or a mixture of any of these. Suitable emulsifyingagents may be, for example, naturally-occurring gums such as gum acaciaor gum tragacanth, naturally-occurring phosphatides such as soya bean,lecithin, esters or partial esters derived from fatty acids and hexitolanhydrides (for example sorbitan monooleate) and condensation productsof the said partial esters with ethylene oxide such as polyoxyethylenesorbitan monooleate. The emulsions may also contain sweetening,flavoring and preservative agents.

Syrups and elixirs may be formulated with sweetening agents such asglycerol, propylene glycol, sorbitol, aspartame or sucrose, and may alsocontain a demulcent, preservative, flavoring and/or coloring agent.

The pharmaceutical compositions may also be in the form of a sterileinjectable aqueous or oily suspension, which may be formulated accordingto known procedures using one or more of the appropriate dispersing orwetting agents and suspending agents, which have been mentioned above.For parenteral formulations, the carrier will usually comprise sterilewater, aqueous sodium chloride solution, 1,3-butanediol, or any othersuitable non-toxic parenterally acceptable diluent or solvent. Otheringredients including those that aid dispersion may be included. Ofcourse, where sterile water is to be used and maintained as sterile, thecompositions and carriers must also be sterilized. Injectablesuspensions may also be prepared, in which case appropriate liquidcarriers, suspending agents and the like may be employed.

Suppository formulations may be prepared by mixing the active ingredientwith a suitable non-irritating excipient that is solid at ordinarytemperatures but liquid at the rectal temperature and will thereforemelt in the rectum to release the drug. Suitable excipients include, forexample, cocoa butter and polyethylene glycols.

Topical formulations, such as creams, ointments, gels and aqueous oroily solutions or suspensions, may generally be obtained by formulatingan active ingredient with a conventional, topically acceptable, vehicleor diluent using conventional procedures well known in the art.

Compositions for administration by insufflation may be in the form of afinely divided powder containing particles of average diameter of, forexample, 30 μm or much less, the powder itself comprising either activeingredient alone or diluted with one or more physiologically acceptablecarriers such as lactose. The powder for insufflation is thenconveniently retained in a capsule containing, for example, 1 to 50 mgof active ingredient for use with a turbo-inhaler device, such as isused for insufflation of the known agent sodium cromoglycate.

Compositions for administration by inhalation may be in the form of aconventional pressurized aerosol arranged to dispense the activeingredient either as an aerosol containing finely divided solid orliquid droplets. Conventional aerosol propellants such as volatilefluorinated hydrocarbons or hydrocarbons may be used and the aerosoldevice is conveniently arranged to dispense a metered quantity of activeingredient.

Compositions for transdermal administration may be in the form of thosetransdermal skin patches that are well known to those of ordinary skillin the art.

Other delivery systems can include time-release, delayed release orsustained release delivery systems. Such systems can avoid repeatedadministrations of the compounds, increasing convenience to the subjectand the physician. Many types of release delivery systems are availableand known to those of ordinary skill in the art. They include polymerbase systems such as poly(lactide-glycolide), copolyoxalates,polycaprolactones, polyesteramides, polyorthoesters, polyhydroxybutyricacid, and polyanhydrides. Microcapsules of the foregoing polymerscontaining drugs are described in, for example, U.S. Pat. No. 5,075,109.Delivery systems also include non-polymer systems that are: lipidsincluding sterols such as cholesterol, cholesterol esters and fattyacids or neutral fats such as mono-di- and tri-glycerides; hydrogelrelease systems; silastic systems; peptide based systems; wax coatings;compressed tablets using conventional binders and excipients; partiallyfused implants; and the like. Specific examples include, but are notlimited to: (a) erosional systems in which an agent of the invention iscontained in a form within a matrix such as those described in U.S. Pat.Nos. 4,452,775, 4,675,189, and 5,736,152, and (b) diffusional systems inwhich an active component permeates at a controlled rate from a polymersuch as described in U.S. Pat. Nos. 3,854,480, 5,133,974 and 5,407,686.In addition, pump-based hardware delivery systems can be used, some ofwhich are adapted for implantation.

For further information on formulations, see Chapter 25.2 in Volume 5 ofComprehensive Medicinal Chemistry (Corwin Hansch; Chairman of EditorialBoard), Pergamon Press 1990, which is specifically incorporated hereinby reference.

The amount of a compound of this invention that is combined with one ormore excipients to produce a single dosage form will necessarily varydepending upon the subject treated, the severity of the disorder orcondition, the rate of administration, the disposition of the compoundand the discretion of the prescribing physician. However, an effectivedosage is in the range of about 0.001 to about 100 mg per kg body weightper day, for example, about 0.5 to about 35 mg/kg/day, in single ordivided doses. For a 70 kg human, this would amount to about 0.0035 to2.5 g/day, such as about 0.05 to about 2.5 g/day. In some instances,dosage levels below the lower limit of the aforesaid range may be morethan adequate, while in other cases still larger doses may be employedwithout causing any harmful side effect, provided that such larger dosesare first divided into several small doses for administration throughoutthe day. For further information on routes of administration and dosageregimes, see Chapter 25.3 in Volume 5 of Comprehensive MedicinalChemistry (Corwin Hansch; Chairman of Editorial Board), Pergamon Press1990, which is specifically incorporated herein by reference.

The size of the dose for therapeutic or prophylactic purposes of acompound of Formula I will naturally vary according to the nature andseverity of the conditions, the age and sex of the animal or patient andthe route of administration, according to well known principles ofmedicine. It will be understood that the specific dosage level andfrequency of dosage for any particular subject may be varied and willdepend upon a variety of factors including the activity of the specificcompound of Formula I, the species, age, body weight, general health,sex and diet of the subject, the mode and time of administration, rateof excretion, drug combination, and severity of the particularcondition, but can nevertheless be routinely determined by one skilledin the art.

A compound of Formula I will in some embodiments be administered to anindividual in combination (e.g., in the same formulation or in separateformulations) with another therapeutic agent (“combination therapy”).The compound of Formula I can be administered in admixture with anothertherapeutic agent or can be administered in a separate formulation. Whenadministered in separate formulations, a compound of Formula I andanother therapeutic agent can be administered substantiallysimultaneously or sequentially.

Such combination treatment may involve, in addition to the compounds ofthe invention, conventional surgery or radiotherapy or chemotherapy.Such chemotherapy may include one or more of the following categories ofanti-tumor agents: (i) antiproliferative/anti-neoplastic drugs andcombinations thereof; (ii) cytostatic agents; (iii) agents which inhibitcancer cell invasion; (iv) inhibitors of growth factor function; (v)antiangiogenic agents; (vi) vascular damaging agents; (vii) antisensetherapies; (viii) gene therapy approaches; (ix) interferon; and (x)immunotherapy approaches.

Therapeutic agents for treating respiratory diseases which may beadministered in combination with a compound of Formula I in a subjectmethod include, but are not limited to beta adrenergics which includebronchodilators including albuterol, isoproterenol sulfate,metaproterenol sulfate, terbutaline sulfate, pirbuterol acetate andsalmeterol formotorol; steroids including beclomethasone dipropionate,flunisolide, fluticasone, budesonide and triamcinolone acetonide.Anti-inflammatory drugs used in connection with the treatment ofrespiratory diseases include steroids such as beclomethasonedipropionate, triamcinolone acetonide, flunisolide and fluticasone.Other anti-inflammatory drugs include cromoglycates such as cromolynsodium. Other respiratory drugs which would qualify as bronchodilatorsinclude anticholenergics including ipratropium bromide. Anti-histaminesinclude, but are not limited to, diphenhydramine, carbinoxamine,clemastine, dimenhydrinate, pryilamine, tripelennamine,chlorpheniramine, brompheniramine, hydroxyzine, cyclizine, meclizine,chlorcyclizine, promethazine, doxylamine, loratadine, and terfenadine.Particular anti-histamines include rhinolast (Astelin®), claratyne(Claritin®), claratyne D (Claritin D®), telfast (Allegra®), Zyrtec®, andbeconase.

In some embodiments, a compound of Formula I is administered as acombination therapy with interferon-gamma (IFN-gamma), a corticosteroidsuch as prednisone, prednisolone, methyl prednisolone, hydrocortisone,cortisone, dexamethasone, betamethasone, etc., or a combination thereof,for the treatment of interstitial lung disease, e.g., idiopathicpulmonary fibrosis.

In some embodiments, a compound of Formula I is administered incombination therapy with a known therapeutic agent used in the treatmentof CF. Therapeutic agents used in the treatment of CF include, but arenot limited to, antibiotics; anti-inflammatory agents; DNAse (e.g.,recombinant human DNAse; pulmozyme; dornase alfa); mucolytic agents(e.g., N-acetylcysteine; Mucomyst™; Mucosil™); decongestants; bronchodilators (e.g., theophylline; ipatropium bromide); and the like.

In another embodiment of the invention, an article of manufacture, or“kit”, containing materials useful for the treatment of the disordersdescribed above is provided. In one embodiment, the kit comprises acontainer comprising a composition of Formula I, or a metabolite,solvate, tautomer, or pharmaceutically acceptable salt or prodrugthereof. In one embodiment, the invention provides a kit for treating aTLR7- and/or TLR8-mediated disorder. In another embodiment, theinvention provides a kit for a condition or disorder treatable byselective modulation of the immune system in a subject. The kit mayfurther comprise a label or package insert on or associated with thecontainer. Suitable containers include, for example, bottles, vials,syringes, blister pack, etc. The container may be formed from a varietyof materials such as glass or plastic. The container holds a compound ofFormula I or a pharmaceutical formulation thereof in an amount effectivefor treating the condition, and may have a sterile access port (forexample, the container may be an intravenous solution bag or a vialhaving a stopper pierceable by a hypodermic injection needle). The labelor package insert indicates that the composition is used for treatingthe condition of choice. In one embodiment, the label or package insertsindicates that the composition comprising a compound of Formula I can beused, for example, to treat a disorder treatable by modulation of TLR7-and/or TLR8-mediated cellular activities. The label or package insertmay also indicate that the composition can be used to treat otherdisorders. Alternatively, or additionally, the kit may further comprisea second container comprising a pharmaceutically acceptable buffer, suchas bacteriostatic water for injection (BWFI), phosphate-buffered saline,Ringer's solution and dextrose solution. It may further include othermaterials desirable from a commercial and user standpoint, includingother buffers, diluents, filters, needles, and syringes.

The kit may further comprise directions for the administration of thecompound of Formula I and, if present, the second pharmaceuticalformulation. For example, if the kit comprises a first compositioncomprising a compound of Formula I and a second pharmaceuticalformulation, the kit may further comprise directions for thesimultaneous, sequential or separate administration of the first andsecond pharmaceutical compositions to a patient in need thereof

In another embodiment, the kits are suitable for the delivery of solidoral forms of a compound of Formula I, such as tablets or capsules. Sucha kit includes, for example, a number of unit dosages. Such kits caninclude a card having the dosages oriented in the order of theirintended use. An example of such a kit is a “blister pack”. Blisterpacks are well known in the packaging industry and are widely used forpackaging pharmaceutical unit dosage forms. If desired, a memory aid canbe provided, for example in the form of numbers, letters, or othermarkings or with a calendar insert, designating the days in thetreatment schedule in which the dosages can be administered.

According to one embodiment, the kit may comprise (a) a first containerwith a compound of Formula I contained therein; and optionally (b) asecond container with a second pharmaceutical formulation containedtherein, wherein the second pharmaceutical formulation comprises asecond compound which may be effective in treating a condition ordisorder treatable by selective modulation of TLR7- and/or TLR8-mediatedcellular activities. Alternatively, or additionally, the kit may furthercomprise a third container comprising a pharmaceutically acceptablebuffer, such as bacteriostatic water for injection (BWFI),phosphate-buffered saline, Ringer's solution and dextrose solution. Itmay further include other materials desirable from a commercial and userstandpoint, including other buffers, diluents, filters, needles, andsyringes.

In certain other embodiments wherein the kit comprises a pharmaceuticalformulation of a compound of Formula I and a second formulationcomprising a second therapeutic agent, the kit may comprise a containerfor containing the separate formulations, such as a divided bottle or adivided foil packet; however, the separate compositions may also becontained within a single, undivided container. Typically, the kitcomprises directions for the administration of the separate components.The kit form is particularly advantageous when the separate componentsare administered in different dosage forms (e.g., oral and parenteral),are administered at different dosage intervals, or when titration of theindividual components of the combination is desired by the prescribingphysician.

EXAMPLES

In order to illustrate the invention, the following examples areincluded. However, it is to be understood that these examples do notlimit the invention and are only meant to suggest a method of practicingthe invention. Persons skilled in the art will recognize that thechemical reactions described may be readily adapted to prepare a numberof other compounds of the invention, and alternative methods forpreparing the compounds of this invention are also deemed to be withinthe scope of this invention. For example, the synthesis ofnon-exemplified compounds according to the invention may be successfullyperformed by modifications apparent to those skilled in the art, e.g.,by appropriately protecting interfering groups, by utilizing othersuitable reagents known in the art other than those described, and/or bymaking routine modifications of reaction conditions. Alternatively,other reactions disclosed herein or known in the art will be recognizedas having applicability for preparing other compounds of the invention.

In the examples described below, unless otherwise indicated alltemperatures are set forth in degrees Celsius. Reagents were purchasedfrom commercial suppliers such as Aldrich Chemical Company, Lancaster,TCI or Maybridge, and were used without further purification unlessotherwise indicated.

The reactions set forth below were done generally under a positivepressure of nitrogen or argon or with a drying tube (unless otherwisestated) in anhydrous solvents, and the reaction flasks were typicallyfitted with rubber septa for the introduction of substrates and reagentsvia syringe. Glassware was oven dried and/or heat dried.

Column chromatography was done on a Biotage system (Manufacturer: DyaxCorporation) having a silica gel column or on a silica SepPak cartridge(Waters). ¹H NMR spectra were recorded on a Varian instrument operatingat 400 MHz. ¹H-NMR spectra were obtained as CDCl₃ solutions (reported inppm), using chloroform as the reference standard (7.25 ppm). When peakmultiplicities are reported, the following abbreviations are used: s(singlet), d (doublet), t (triplet), m (multiplet), br (broadened), dd(doublet of doublets), dt (doublet of triplets). Coupling constants,when given, are reported in Hertz (Hz).

Example 1 Synthesis of (1E,4E)-ethyl2-amino-8-(perfluoroethyl)-3H-benzo[b]azepine-4-carboxylate (7)

A reaction scheme for the synthesis of compound (7) is shown in FIG. 1.

Step A: Preparation of potassium 2,2,3,3,3-pentafluoropropanoate:Pentafluoropropionic acid (20.5 g, 183 mmol) was slowly added to asolution of KOtBu (29.9 g, 183 mmol) in ether (400 mL) at 0° C. After 30minutes the ice bath was removed. After stirring at room temperature for4 hours the suspension was filtered and the cake was washed with ether(200 mL). The fine white solid was placed under vacuum for 16 hoursprior to use.

Step B: Preparation of 1-methyl-4-(perfluoroethyl)benzene (2): Procedureadapted from Syn. Comm., (1988) 18(9):965-972. A mixture of copperiodide (129 g, 679 mmol), 1-iodo-4-methylbenzene (1) (74 g, 339 mmol)and potassium 2,2,3,3,3-pentafluoropropanoate (120 g, 594 mmol) weredissolved in DMF (500 mL) and heated to 120° C. for 30 minutes and thenheated at 160° C. for 4 hours, after which the mixture was allowed tocool to room temperature. Water (200 mL) and ether (200 mL) were addedand after stirring for 30 minutes the solids were filtered and washedwith ether. The phases were separated and the organic layer was washedwith brine/water (3×250 mL). The organic layer was dried andconcentrated to yield 1-methyl-4-(perfluoroethyl)benzene (66 g, 92%) asa dark oil.

Step C: Preparation of 1-methyl-2-nitro-4-(perfluoroethyl)benzene (3):Prepared from 1-methyl-4-(perfluoroethyl)benzene (1.95 g, 9.28 mmol)with sulfuric and nitric acid as described in EP 0418175 (1.45 g, 61%).

Step D: Preparation of(E)-N,N-dimethyl-2-(2-nitro-4-(perfluoroethyl)phenyl)ethenamine (4): Toa solution of 1-methyl-2-nitro-4-(perfluoroethyl)benzene (10.0 g, 39.2mmol) and pyrollidine (2.79 g, 39.2 mmol) in toluene (250 mL) was addeddimethyl formamide dimethyl acetal (4.96 g, 39.2 mmol). The mixture washeated to reflux for 16 hours. After cooling to room temperature themixture was concentrated under vacuum and the resulting oil wasimmediately used in the next reaction.

Step E: Preparation of 2-nitro-4-(perfluoroethyl)benzaldehyde (5): To asolution of crude(E)-N,N-dimethyl-2-(2-nitro-4-(perfluoroethyl)phenyl)ethenamine (12.2 g,39.2 mmol) in THF (300 mL) and pH 7.2 phosphate buffer (300 mL) wasadded NaIO₄ (29.3 g, 137.2 mmol). After 2.5 hours an aliquot (−0.3 mL)was removed, filtered and dried. The proton NMR of this sample indicatedthat the reaction was complete. The solids were removed and the filtercake was washed with EtOAc. The filtrate was washed with brine (2×100mL), dried and concentrated. The concentrate was purified via flashchromatography (100% hexane to 5% EtOAc) to provide2-nitro-4-(perfluoroethyl)benzaldehyde (5.4 g, 52%).

Step F: Preparation of (E)-ethyl2-(cyanomethyl)-3-(2-nitro-4-(perfluoroethyl)phenyl)acrylate (6): To asolution of 2-nitro-4-(perfluoroethyl)benzaldehyde (5.4 g, 20.1 mmol) intoluene (150 mL) was added α-cyanomethylcarboethoxyethylidenetriphenylphosphorane (8.55 g, 22.1 mmol). The mixture was heated to 75°C. for 30 minutes. The reaction was allowed to cool and the solvent wasremoved under vacuum. The concentrate was purified via flashchromatography (100% hexanes to 20% EtOAc) to provide (E)-ethyl2-(cyanomethyl)-3-(2-nitro-4-(perfluoroethyl)phenyl)acrylate (6.00 g,79%).

Step G: Preparation of (1E,4E)-ethyl2-amino-8-(perfluoroethyl)-3H-benzo[b]azepine-4-carboxylate (7): To asolution of (E)-ethyl2-(cyanomethyl)-3-(2-nitro-4-(perfluoroethyl)phenyl)acrylate (2.60 g,6.87 mmol) was added iron powder (2.30 g, 41.2 mmol). The mixture washeated to 90° C. for 5 hours. Upon cooling the acetic acid was removedunder vacuum and the resulting semisolid was dissolved in 50% K₂CO₃ (100mL) and EtOAc (100 mL). The mixture was filtered to remove insolublematerial and the phases were separated. The aqueous phase was extractedwith EtOAc (2×100 mL). The combined organics were dried andconcentrated. The concentrate was purified via flash chromatography(100% CH₂Cl₂ to 2% MeOH) to yield (1E,4E)-ethyl2-amino-8-(perfluoroethyl)-3H-benzo[b]azepine-4-carboxylate (1.8 g,74%). ¹H NMR (CDCl₃) δ 1.39 (t, 3H), 2.95 (s, 2H), 4.32 (q, 2H), 5.12(br s, 1-2H), 7.22-7.27 (m, 2H), 7.47-7.51 (m, 2H), 7.80 (s, 1H).

FIG. 2 shows a reaction scheme for the synthesis of compound of thegeneral formula (8). The following amides were prepared as shown in FIG.2.

Example 2 Synthesis of(1E,4E)-2-amino-N,N-bis(2-methoxyethyl)-8-(perfluoroethyl)-3H-benzo[b]azepine-4-carboxamide(9)

Trimethylaluminum (0.34 mL of a 2.0 M solution in toluene) was added tobis(2-methoxyethyl)amine (92 mg, 0.69 mmol) in DCE (3 mL). After 10minutes solid (1E,4E)-ethyl2-amino-8-(perfluoroethyl)-3H-benzo[b]azepine-4-carboxylate (80 mg, 0.23mmol) was added and the vessel was sealed and heated to 75° C. for 16-20hours. Upon cooling the reaction was quenched with saturated Rochelle'ssalt (2 mL) and after 20 minutes the mixture was partitioned betweenCH₂Cl₂ (50 mL) and brine (50 mL). The phases were separated and theaqueous was extracted with CH₂Cl₂ (2×20 mL). The combined organics weredried and concentrated. The crude material was purified via preparativeTLC (2, 0.5 mm plates, eluting with 5-10% MeOH/CH₂Cl₂ with 4-6 drops ofNH₄OH). ¹H NMR (CDCl₃) δ 2.81 (s, 2H), 3.36 (s, 6H), 3.55-3.74 (m, 8H),6.98 (s, 1H), 7.20 (d, 1H), 7.40 (d, 1H), 7.39 (s, 1H).

Example 3 Synthesis of(1E,4E)-2-amino-N,N-diethyl-8-(perfluoroethyl)-3H-benzo[b]azepine-4-carboxamide(10)

Compound (9) was prepared as described in Example 2 to provide 17 mg(21%) of the desired product. ¹H NMR (CDCl₃) δ 1.22-1.27 (m, 6H), 1.78(s, 2H), 3.49-3.56 (m, 4H), 5.04 (br s, 1h), 6.98 (s, 1H), 7.20 (d, 1H),7.38 (d, 1H), 7.48 (s, 1H).

Example 4 Synthesis of(1E,4E)-2-amino-8-(perfluoroethyl)-N,N-dipropyl-3H-benzo[b]azepine-4-carboxamide(11)

Compound (10) was prepared as described in Example 2 to provide 24 mg(28%) of the desired product. ¹H NMR (CDCl₃) δ 0.85-0.94 (m, 6H),1.61-1.71 (m, 4H), 2.77 (s, 2H), 3.40-3.55 (m, 4H), 6.81 (s, 1H), 7.21(d, 1H), 7.37 (d, 1H), 7.48 (s, 1H).

FIG. 3 shows a reaction scheme for the synthesis of compounds of thegeneral formula (12). The following amides were prepared as shown inFIG. 3.

Example 5 Synthesis of(1E,4E)-2-amino-N-ethyl-8-(perfluoroethyl)-3H-benzo[b]azepine-4-carboxamide(15)

Step A: Preparation of (1E,4E)-ethyl2-(tert-butoxycarbonyl)-8-(perfluoroethyl)-3H-benzo[b]azepine-4-carboxylate(13): To a solution of (1E,4E)-ethyl2-amino-8-(perfluoroethyl)-3H-benzo[b]azepine-4-carboxylate (7) (2.28 g,6.55 mmol) in CH₂Cl₂ (150 mL) was added Boc anhydride (1.43 g, 6.55mmol). After 24 hours the mixture was concentrated and purified viaflash chromatography (100% hexanes to 10% EtOAc) to yield (1E,4E)-ethyl2-(tert-butoxycarbonyl)-8-(perfluoroethyl)-3H-benzo[b]azepine-4-carboxylate(1.94 g, 66%).

Step B: Preparation of(1E,4E)-2-(tert-butoxycarbonyl)-8-(perfluoroethyl)-3H-benzo[b]azepine-4-carboxylicacid (14): To a solution of (1E,4E)-ethyl2-(tert-butoxycarbonyl)-8-(perfluoroethyl)-3H-benzo[b]azepine-4-carboxylatein THF/H₂O (4:1, 10 mL) was added LiOH.H₂O (42 mg, 1.00 mmol). After 4hours the mixture was made acidic (pH 4) with 10% citric acid andextracted with CH₂Cl₂ (2×100 mL). The combined organics were dried andconcentrated. The material was used without further purification.

Step C: Preparation of(1E,4E)-2-amino-N-ethyl-8-(perfluoroethyl)-3H-benzo[b]azepine-4-carboxamide(15): To a solution of triethylamine (76 mg, 0.750 mmol), PyBOP (143 mg,0.275 mmol) and(1E,4E)-2-(tert-butoxycarbonyl)-8-(perfluoroethyl)-3H-benzo[b]azepine-4-carboxylicacid (105 mg, 0.250 mmol) in DMF (4 mL) was added ethylaminehydrochloride (41 mg, 0.500 mmol). After stirring for 14 hours themixture was partitioned between EtOAc (50 mL) and brine (100 mL). Thephases were separated and the organic layer was washed with water (3×100mL). The combined organics were dried and concentrated. The cruderesidue was taken up in CH₂Cl₂/TFA (˜1:1, 50 mL) and stirred at roomtemperature for 2 hours. The volatiles were removed under vacuum and theresidue was partitioned between EtOAc and 10% Na₂CO₃ (1:1, 100 mL). Thephases were separated and the aqueous was extracted with EtOAc (2×50mL). The combined organics were dried and concentrated. The concentratewas purified via preparative TLC (2, 0.5 mm plates eluting with 10%MeOH/CH₂Cl₂ with 4-6 drops of NH₄OH) to provide(1E,4E)-2-amino-N-ethyl-8-(perfluoroethyl)-3H-benzo[b]azepine-4-carboxamide(6.5 mg, 8%). ¹H NMR (CDCl₃) δ 1.26 (t, 2H), 2.94 (s, 2H), 3.41-3.48 (m,2H), 5.15 (br s, 1H), 6.08 (br s, 1H), 7.16 (s, 1H), 7.22 (d, 1H), 7.42(d, 1H), 7.47 (s, 1H).

Example 6 Synthesis of(1E,4E)-2-amino-8-(perfluoroethyl)-N-propyl-3H-benzo[b]azepine-4-carboxamide(16)

Compound (16) was prepared as described in Example 5, substitutingpropylamine for ethylamine hydrochloride, to provide 16 mg (45%) of thedesired product.

¹H NMR (CDCl₃) δ 0.99 (s, 3H), 1.59-1.68 (m, 2H), 2.94 (s, 2H),3.34-3.92 (m, 2H), 5.19 (br s, 1H), 6.14 (br s, 1H), 7.15 (s, 1H), 7.22(d, 1H), 7.42 (d, 1H), 7.48 (s, 1H).

Example 7 Synthesis of (1E,4E)-ethyl2-amino-8-(pyrrolidine-1-carbonyl)-3H-benzo[b]azepine-4-carboxylate (24)

The reaction scheme for the synthesis of compound (24) is shown in FIG.4.

Step A: Preparation of(E)-2-(4-bromo-2-nitrophenyl)-N,N-dimethylethenamine (18): To a solutionof 1-methyl-2-nitro-4-bromobenzene (17) (29.86 g, 138.2 mmol) in toluene(200 mL) was added dimethylformamide dimethylacetal (17.52 g, 138.2mmol). The mixture was heated to reflux for 14 hours. After cooling toroom temperature the mixture was concentrated under vacuum and theresulting oil was immediately used in the next reaction.

Step B: Preparation of 4-bromo-2-nitrobenzaldehyde (19): To a solutionof crude (E)-2-(4-bromo-2-nitrophenyl)-N,N-dimethylethenamine (35.5 g,131 mmol) in THF (300 mL) and pH 7.2 phosphate buffer (300 mL) was addedNaIO₄ (56.0 g, 262 mmol). The solids were removed and the filter cakewas washed with EtOAc (200 mL). The filtrate was washed with brine(2×100 mL), dried and concentrated. The concentrate was purified viaflash chromatography (5% EtOAc/hexanes to 10% EtOAc/hexanes) to provide4-bromo-2-nitrobenzaldehyde (8.41 g, 28% yield).

Step C: Preparation of (E)-ethyl3-(4-bromo-2-nitrophenyl)-2-(cyanomethyl)acrylate (20): To a solution of4-bromo-2-nitrobenzaldehyde (3.45 g, 15.0 mmol) in toluene (15 mL) wasadded α-cyanomethylcarboethoxyethylidene triphenylphosphorane (6.10 g,15.7 mmol). The mixture was heated to 75° C. for 16 hours. The reactionwas allowed to cool and the solvent was removed under vacuum. Theconcentrate was purified via flash chromatography (100% hexanes to 20%EtOAc) to yield (E)-ethyl3-(4-bromo-2-nitrophenyl)-2-(cyanomethyl)acrylate (2.25 g, 44% yield) asan off white solid.

Step D: Preparation of (1E,4E)-ethyl2-amino-8-bromo-3H-benzo[b]azepine-4-carboxylate (21): To a solution of(E)-ethyl 3-(4-bromo-2-nitrophenyl)-2-(cyanomethyl)acrylate (1.00 g, 2.9mmol) in acetic acid (25 mL) was added iron powder (1.10 g, 19.0 mmol).The mixture was heated to 90° C. for 5 hours. Upon cooling the aceticacid was removed under vacuum and the resulting semisolid was dissolvedin 50% K₂CO₃ (100 mL) and EtOAc (100 mL). The mixture was filtered toremove insoluble material and the phases were separated. The aqueousphase was extracted with EtOAc (2×100 mL). The combined organics weredried and concentrated. The concentrate was purified via flashchromatography (Biotage 40m, 5% MeOH/CH₂Cl₂) to yield (1E,4E)-ethyl2-amino-8-bromo-3H-benzo[b]azepine-4-carboxylate (0.52 g, 57%).

Step E: Preparation of(1E,4E)-ethyl-8-bromo-2-(tert-butoxycarbonyl)-3H-benzo[b]azepine-4-carboxylate(22): To a CH₂Cl₂ (5 mL) solution containing (1E,4E)-ethyl2-amino-8-bromo-3H-benzo[b]azepine-4-carboxylate (198 mg, 0.640 mmol)was added Boc anhydride (140 mg, 0.640 mmol). The solution was stirredat room temperature for 72 hours. The reaction was concentrated todryness and purified by column chromatography (Biotage 12m, 4:1hexanes:EtOAc) to provide(1E,4E)-ethyl-8-bromo-2-(tert-butoxycarbonyl)-3H-benzo[b]azepine-4-carboxylate(245 mg, 94% yield) as a white solid.

Step F: Preparation of(1E,4E)-ethyl-2-(tert-butoxycarbonyl)-8-(pyrrolidine-1-carbonyl)-3H-benzo[b]azepine-4-carboxylate(23): To an ethanol solution (15 mL) containing K₃PO₄ (938 mg, 4.42mmol), 4-(pyrrolidine-1-carbonyl)phenylboronic acid (785 mg, 3.58 mmol),and(1E,4E)-ethyl-8-bromo-2-(tert-butoxycarbonyl)-3H-benzo[b]azepine-4-carboxylate(489 mg, 1.19 mmol), was added palladium acetate (80.5 mg, 0.358 mmol).The reaction was heated to 60° C. for 2 hours, then cooled to roomtemperature and concentrated to dryness. The brown oil was purified bypreparative LC plate (100% EtOAc) to provide(1E,4E)-ethyl-2-(tert-butoxycarbonyl)-8-(pyrrolidine-1-carbonyl)-3H-benzo[b]azepine-4-carboxylate(277 mg, 46% yield) as a tan oil.

Step G: Preparation of (1E,4E)-ethyl2-amino-8-(pyrrolidine-1-carbonyl)-3H-benzo[b]azepine-4-carboxylate(24):(1E,4E)-ethyl-2-(tert-butoxycarbonyl)-8-(pyrrolidine-1-carbonyl)-3H-benzo[b]azepine-4-carboxylate(110 mg, 0.218 mmol) was diluted with a 1:4 TFA:CH₂Cl₂ solution (4 mL).The reaction was stirred at room temperature for 1 hour, and thendiluted with CH₂Cl₂. The organic phase was washed with 10% K₂CO₃ andbrine (30 mL). The CH₂Cl₂ solution was dried over Na₂SO₄, filtered, andconcentrated to provide (1E, 4E)-ethyl2-amino-8-(pyrrolidine-1-carbonyl)-3H-benzo[b]azepine-4-carboxylate (88mg, 81% yield) as a yellow solid. ¹H NMR (CDCl₃) δ 1.39 (t, 3H),1.88-1.99 (m, 4H), 2.98 (s, 2H), 3.49-3.52 (m, 2H), 3.66-3.69 (m, 2H),4.30-4.35 (m, 2H), 7.32 (d, 1H), 7.46-7.49 (m, 2H), 7.60 (d, 2H) 7.67(d, 2H), 7.84 (s, 1H).

Example 8 Synthesis of (1E,4E)-ethyl2-amino-8-(4-(methoxycarbonyl)phenyl)-3H-benzo[b]azepine-4-carboxylate(25)

Compound (25) was prepared according to the general procedures describedin Example 7, substituting 4-(methoxycarbonyl)phenylboronic acid for4-(pyrrolidine-1-carbonyl)phenylboronic acid, to provide (1E,4E)-ethyl2-amino-8-(4-(methoxycarbonyl)phenyl)-3H-benzo[b]azepine-4-carboxylate(17 mg, 0.031 mmol, 54% yield) as a yellow solid. ¹H NMR (CDCl₃) δ 1.39(t, 3H), 2.99 (s, 2H), 3.94 (s, 3H), 4.32 (q, 2H), 7.33 (dd, 1H),7.46-7.49 (m, 2H), 7.71 (dd, 2H), 7.83 (s, 1H), 8.09 (s, 1H), 8.11 (s,1H).

Example 9 Synthesis of (1E,4E)-ethyl2-amino-8-(4-(methylcarbamoyl)phenyl)-3H-benzo[b]azepine-4-carboxylate(26)

Compound (26) was prepared according to the general procedures describedin Example 7, substituting 4-(methylcarbamoyl)phenylboronic acid for4-(pyrrolidine-1-carbonyl)phenylboronic acid, to provide (1E,4E)-ethyl2-amino-8-(4-(methoxycarbonyl)phenyl)-3H-benzo[b]azepine-4-carboxylate(1 mg, 0.031 mmol, 21% yield) as a yellow solid. ¹H NMR (CDCl₃) δ 1.39(t, 3H), 2.98 (s, 2H), 3.06 (s, 3H), 4.31-4.36 (q, 2H), 7.33 (dd, 1H),7.46-7.49 (m, 2H), 7.34 (d, 1H), 7.47-7.52 (m, 3H), 7.32 (d, 2H)7.83-7.85 (m, 3H).

Example 10 Synthesis of(1E,4E)-2-amino-N,N-dipropyl-8-(4-(pyrrolidine-1-carbonyl)phenyl)-3H-benzo[b]azepine-4-carboxamide(27)

Compound (27) was prepared from compound (24) by a method similar tothat described in Example 2 to provide 49 mg (43%) of the desiredcompound. ¹H NMR (CDCl₃) δ 0.93 (t, 6H), 1.63-1.71 (m, 4H), 1.89 (m,2H), 1.98 (m, 2H), 2.83 (s, 2H), 3.40-3.51 (m, 6H), 3.67 (t, 2H), 6.83(s, 1H), 7.3 (dd, 1H), 7.35 (d, 1H), 7.49 (d, 1H), 7.64 (q, 4H).

The activity of the compounds of this invention may be determined by thefollowing assays.

Example 11 HEK/TLR Assays

Human embryonic kidney (HEK) cells which stably express various humanTLR genes, including TLR7 and TLR8, and a NFkB-luciferase reporter genewere incubated with various concentrations of compound overnight. Theamount of induced luciferase was measured by reading the absorbance at650 nm. Compounds of this invention have an MC₅₀ of 100 μM or less,wherein MC₅₀ is defined as the concentration at which 50% of maximuminduction is seen.

Example 12 PBMC Assays for TLR7 and TLR8

Peripheral blood mononuclear cells (PBMCs) from human blood wereisolated using BD Vacutainer Cell Preparation Tubes with sodium citrate.Cells were incubated with compound overnight. TLR8 activity was assayedby measuring the amount of TNFα in supernatants by ELISA. TLR7 activitywas assayed by measuring the amount of IFNα in supernatants by ELISA(R&D Systems). Compounds of this invention had an MC₅₀ of 100 μM orless, wherein MC₅₀ is the concentration at which 50% of the maximuminduction is seen.

The foregoing description is considered as illustrative only of theprinciples of the invention. Further, since numerous modifications andchanges will be readily apparent to those skilled in the art, it is notdesired to limit the invention to the exact construction and processshown as described above. Accordingly, all suitable modifications andequivalents may be resorted to falling within the scope of the inventionas defined by the claims that follow.

The words “comprise,” “comprising,” “include,” “including,” and“includes” when used in this specification and in the following claimsare intended to specify the presence of stated features, integers,components, or steps, but they do not preclude the presence or additionof one or more other features, integers, components, steps, or groupsthereof.

1. A method of treating cancer, comprising administering to a mammal inneed thereof an effective amount of a compound of the Formula

a tautomer, or a pharmaceutically acceptable salt thereof, wherein: saidcancer is selected from biliary tract cancer, brain cancer, breastcancer, cervical cancer, choriocarcinoma, colon cancer, endometrialcancer, esophageal cancer, gastric cancer, intraepithelial neoplasms,leukemia, lymphoma, liver cancer, lung cancer, melanoma, neuroblastomas,oral cancer, ovarian cancer, pancreatic cancer, prostate cancer, rectalcancer, renal cancer, sarcomas, skin cancer, testicular cancer, thyroidcancer, other carcinomas and sarcomas; Y is an aryl or heteroaryl ring,wherein said aryl and heteroaryl rings are substituted with one or moregroups independently selected from alkenyl, alkynyl, Br, CN, OH, NR⁶R⁷,C(═O)R⁸, NR⁶SO₂R⁷, (C₁-C₆ alkyl)amino, R⁶OC(═O)CH═CH—, SR⁶ and SO₂R⁶,and wherein said aryl and heteroaryl rings are optionally furthersubstituted with one or more groups independently selected from F, Cl,CF₃, CF₃O—, HCF₂O—, alkyl, heteroalkyl and ArO—; R¹, R³ and R⁴ areindependently selected from H, alkyl, alkenyl, alkynyl, heteroalkyl,cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl and heteroaryl, whereinsaid alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, cycloalkenyl,heterocycloalkyl, aryl and heteroaryl are optionally substituted withone or more groups independently selected from alkyl, alkenyl, alkynyl,F, Cl, Br, I, CN, OR⁶, NR⁶R⁷, C(═O)R⁶, C(═O)OR⁶, OC(═O)R⁶, C(═O)NR⁶R⁷,(C₁-C₆ alkyl)amino, CH₃OCH₂O—, R⁶OC(═O)CH═CH—, NR⁶SO₂R⁷, SR⁶ and SO₂R⁶,or R³ and R⁴ together with the atom to which they are attached form asaturated or partially unsaturated carbocyclic ring, wherein saidcarbocyclic ring is optionally substituted with one or more groupsindependently selected from alkyl, alkenyl, alkynyl, F, Cl, Br, I, CN,OR⁶, NR⁶R⁷, C(═O)R⁶, C(═O)OR⁶, OC(═O)R⁶, C(═O)NR⁶R⁷, (C₁-C₆ alkyl)amino,CH₃OCH₂O—, R⁶OC(═O)CH═CH—, NR⁶SO₂R⁷, SR⁶ and SO₂R⁶; R² and R⁸ areindependently selected from H, OR⁶, NR⁶R⁷, alkyl, alkenyl, alkynyl,heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl andheteroaryl, wherein said alkyl, alkenyl, alkynyl, heteroalkyl,cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl and heteroaryl areoptionally substituted with one or more groups independently selectedfrom alkyl, alkenyl, alkynyl, F, Cl, Br, I, CN, OR⁶, NR⁶R⁷, C(═O)R⁶,C(═O)OR⁶, OC(═O)R⁶, C(═O)NR⁶R⁷, (C₁-C₆ alkyl)amino, CH₃OCH₂O—,R⁶OC(═O)CH═CH—, NR⁶SO₂R⁷, SR⁶ and SO₂R⁶; R^(5a), R^(5b), and R^(5c) areindependently H, F, Cl, Br, I, OMe, CH₃, CH₂F, CHF₂ or CF₃; and R⁶ andR⁷ are independently H, alkyl, or heteroalkyl, such that said cancer istreated.
 2. The method of claim 1, wherein said cancer is colon cancer,ovarian cancer, breast cancer, or lymphoma.
 3. The method of claim 1,wherein R² is OR⁶.
 4. The method of claim 3, wherein R⁶ is alkyl.
 5. Themethod of claim 4, wherein R⁶ is ethyl.
 6. The method of claim 1,wherein R² is NR⁶R⁷.
 7. The method of claim 1, wherein R⁶ and R⁷ areindependently H, ethyl, propyl, or CH₂CH₂OCH₃.
 8. The method of claim 1,wherein Y is aryl.
 9. The method of claim 8, wherein said aryl issubstituted with C(═O)R⁸.
 10. The method of claim 9, wherein R⁸ is OR⁶,NR⁶R⁷ or heterocycloalkyl.
 11. The method of claim 10, wherein R⁶ and R⁷are independently selected from H and alkyl.
 12. The method of claim 1,wherein Y is


13. The method of claim 1, wherein R¹, R³, R⁴, R^(5a), R^(5b) and R^(5c)are each hydrogen.
 14. The method of claim 1, wherein said compound isselected from: (1E,4E)-ethyl2-amino-8-(4-(pyrrolidine-1-carbonyl)phenyl)-3H-benzo[b]azepine-4-carboxylate;(1E,4E)-ethyl2-amino-8-(4-(methoxycarbonyl)phenyl)-3H-benzo[b]azepine-4-carboxylate;(1E,4E)-ethyl2-amino-8-(4-(methylcarbamoyl)phenyl)-3H-benzo[b]azepine-4-carboxylate;(1E,4E)-2-amino-N,N-dipropyl-8-(4-(pyrrolidine-1-carbonyl)phenyl)-3H-benzo[b]azepine-4-carboxamide;and pharmaceutically acceptable salts thereof.
 15. The method of claim1, wherein said effective amount is about 0.5 to about 35 mg/kg/day. 16.A method of treating allergy, comprising administering to a mammal inneed thereof an effective amount of a compound of the Formula

a tautomer, or a pharmaceutically acceptable salt thereof, wherein: Y isan aryl or heteroaryl ring, wherein said aryl and heteroaryl rings aresubstituted with one or more groups independently selected from alkenyl,alkynyl, Br, CN, OH, NR⁶R⁷, C(═O)R⁸, NR⁶SO₂R⁷, (C₁-C₆ alkyl)amino,R⁶OC(═O)CH═CH—, SR⁶ and SO₂R⁶, and wherein said aryl and heteroarylrings are optionally further substituted with one or more groupsindependently selected from F, Cl, CF₃, CF₃O—, HCF₂O—, alkyl,heteroalkyl and ArO—; R¹, R³ and R⁴ are independently selected from H,alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, cycloalkenyl,heterocycloalkyl, aryl and heteroaryl, wherein said alkyl, alkenyl,alkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, aryland heteroaryl are optionally substituted with one or more groupsindependently selected from alkyl, alkenyl, alkynyl, F, Cl, Br, I, CN,OR⁶, NR⁶R⁷, C(═O)R⁶, C(═O)OR⁶, OC(═O)R⁶, C(═O)NR⁶R⁷, (C₁-C₆ alkyl)amino,CH₃OCH₂O—, R⁶OC(═O)CH═CH—, NR⁶SO₂R⁷, SR⁶ and SO₂R⁶, or R³ and R⁴together with the atom to which they are attached form a saturated orpartially unsaturated carbocyclic ring, wherein said carbocyclic ring isoptionally substituted with one or more groups independently selectedfrom alkyl, alkenyl, alkynyl, F, Cl, Br, I, CN, OR⁶, NR⁶R⁷, C(═O)R⁶,C(═O)OR⁶, OC(═O)R⁶, C(═O)NR⁶R⁷, (C₁-C₆ alkyl)amino, CH₃OCH₂O—,R⁶OC(═O)CH═CH—, NR⁶SO₂R⁷, SR⁶ and SO₂R⁶; R² and R⁸ are independentlyselected from H, OR⁶, NR⁶R⁷, alkyl, alkenyl, alkynyl, heteroalkyl,cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl and heteroaryl, whereinsaid alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, cycloalkenyl,heterocycloalkyl, aryl and heteroaryl are optionally substituted withone or more groups independently selected from alkyl, alkenyl, alkynyl,F, Cl, Br, I, CN, OR⁶, NR⁶R⁷, C(═O)R⁶, C(═O)OR⁶, OC(═O)R⁶, C(═O)NR⁶R⁷,(C₁-C₆ alkyl)amino, CH₃OCH₂O—, R⁶OC(═O)CH═CH—, NR⁶SO₂R⁷, SR⁶ and SO₂R⁶;R^(5a), R^(5b), and R^(5c) are independently H, F, Cl, Br, I, OMe, CH₃,CH₂F, CHF₂ or CF₃; and R⁶ and R⁷ are independently H, alkyl, orheteroalkyl, such that said allergy is treated.
 17. The method of claim16, wherein said allergy is acquired hypersensitivity to an allergen.18. The method of claim 17, wherein said allergy is selected fromeczema, allergic rhinitis or coryza, hay fever, asthma, urticaria(hives) and food allergies, and other atopic conditions.
 19. The methodof claim 16, wherein said compound is selected from: (1E,4E)-ethyl2-amino-8-(4-(pyrrolidine-1-carbonyl)phenyl)-3H-benzo[b]azepine-4-carboxylate;(1E,4E)-ethyl2-amino-8-(4-(methoxycarbonyl)phenyl)-3H-benzo[b]azepine-4-carboxylate;(1E,4E)-ethyl2-amino-8-(4-(methylcarbamoyl)phenyl)-3H-benzo[b]azepine-4-carboxylate;(1E,4E)-2-amino-N,N-dipropyl-8-(4-(pyrrolidine-1-carbonyl)phenyl)-3H-benzo[b]azepine-4-carboxamide;and pharmaceutically acceptable salts thereof.
 20. The method of claim16, wherein said effective amount is about 0.5 to about 35 mg/kg/day.